CN105214077B - Application of the USP33 in tumour - Google Patents
Application of the USP33 in tumour Download PDFInfo
- Publication number
- CN105214077B CN105214077B CN201410242697.3A CN201410242697A CN105214077B CN 105214077 B CN105214077 B CN 105214077B CN 201410242697 A CN201410242697 A CN 201410242697A CN 105214077 B CN105214077 B CN 105214077B
- Authority
- CN
- China
- Prior art keywords
- usp33
- cell
- lung cancer
- expression
- tumour
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 101000748157 Homo sapiens Ubiquitin carboxyl-terminal hydrolase 33 Proteins 0.000 title claims abstract description 135
- 102100040047 Ubiquitin carboxyl-terminal hydrolase 33 Human genes 0.000 title claims abstract description 135
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 56
- 238000004393 prognosis Methods 0.000 claims abstract description 15
- 238000001514 detection method Methods 0.000 claims abstract description 13
- 239000003814 drug Substances 0.000 claims abstract description 7
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 89
- 201000005202 lung cancer Diseases 0.000 claims description 67
- 208000020816 lung neoplasm Diseases 0.000 claims description 67
- 108700025716 Tumor Suppressor Genes Proteins 0.000 claims description 6
- 102000044209 Tumor Suppressor Genes Human genes 0.000 claims description 6
- 208000031261 Acute myeloid leukaemia Diseases 0.000 claims description 5
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 claims description 4
- 230000033228 biological regulation Effects 0.000 claims description 3
- 238000011161 development Methods 0.000 claims description 3
- 229940079593 drug Drugs 0.000 claims description 3
- 201000001441 melanoma Diseases 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 230000014509 gene expression Effects 0.000 abstract description 53
- 238000000034 method Methods 0.000 abstract description 11
- 239000003550 marker Substances 0.000 abstract description 5
- 208000001894 Nasopharyngeal Neoplasms Diseases 0.000 abstract description 2
- 230000007423 decrease Effects 0.000 abstract description 2
- 230000001105 regulatory effect Effects 0.000 abstract description 2
- 230000001276 controlling effect Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 74
- 101150097792 Robo1 gene Proteins 0.000 description 28
- 239000000523 sample Substances 0.000 description 26
- 201000005296 lung carcinoma Diseases 0.000 description 22
- 210000001519 tissue Anatomy 0.000 description 17
- 108020004459 Small interfering RNA Proteins 0.000 description 14
- 201000011510 cancer Diseases 0.000 description 14
- 210000004072 lung Anatomy 0.000 description 13
- 238000004458 analytical method Methods 0.000 description 11
- 230000035772 mutation Effects 0.000 description 11
- 101150087227 usp33 gene Proteins 0.000 description 11
- 108090000623 proteins and genes Proteins 0.000 description 10
- 230000000694 effects Effects 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 9
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 9
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 9
- 206010006187 Breast cancer Diseases 0.000 description 8
- 208000026310 Breast neoplasm Diseases 0.000 description 8
- YPHMISFOHDHNIV-FSZOTQKASA-N cycloheximide Chemical compound C1[C@@H](C)C[C@H](C)C(=O)[C@@H]1[C@H](O)CC1CC(=O)NC(=O)C1 YPHMISFOHDHNIV-FSZOTQKASA-N 0.000 description 8
- 238000011160 research Methods 0.000 description 8
- 108700002783 roundabout Proteins 0.000 description 8
- 230000012292 cell migration Effects 0.000 description 7
- 238000012546 transfer Methods 0.000 description 7
- 238000001262 western blot Methods 0.000 description 7
- 108090000848 Ubiquitin Proteins 0.000 description 6
- 102000044159 Ubiquitin Human genes 0.000 description 6
- 230000005012 migration Effects 0.000 description 6
- 238000013508 migration Methods 0.000 description 6
- 238000012545 processing Methods 0.000 description 6
- 210000004881 tumor cell Anatomy 0.000 description 6
- 108700011893 Slit homolog 2 Proteins 0.000 description 5
- 102100027340 Slit homolog 2 protein Human genes 0.000 description 5
- 101150085024 Slit2 gene Proteins 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 5
- 238000010186 staining Methods 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 4
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 238000000749 co-immunoprecipitation Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 230000007246 mechanism Effects 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 238000001890 transfection Methods 0.000 description 4
- 230000029663 wound healing Effects 0.000 description 4
- 208000010507 Adenocarcinoma of Lung Diseases 0.000 description 3
- 108700019961 Neoplasm Genes Proteins 0.000 description 3
- 102000048850 Neoplasm Genes Human genes 0.000 description 3
- 206010041067 Small cell lung cancer Diseases 0.000 description 3
- 206010052428 Wound Diseases 0.000 description 3
- 208000027418 Wounds and injury Diseases 0.000 description 3
- 230000003197 catalytic effect Effects 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 230000009504 deubiquitination Effects 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 230000002255 enzymatic effect Effects 0.000 description 3
- 210000004907 gland Anatomy 0.000 description 3
- 238000011532 immunohistochemical staining Methods 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 230000009545 invasion Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 230000001404 mediated effect Effects 0.000 description 3
- 108020004999 messenger RNA Proteins 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000019491 signal transduction Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- 206010069754 Acquired gene mutation Diseases 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- 238000009007 Diagnostic Kit Methods 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 2
- 101000607909 Homo sapiens Ubiquitin carboxyl-terminal hydrolase 1 Proteins 0.000 description 2
- 101000777120 Homo sapiens Ubiquitin carboxyl-terminal hydrolase 44 Proteins 0.000 description 2
- 101000944219 Homo sapiens cAMP-dependent protein kinase catalytic subunit beta Proteins 0.000 description 2
- 102100038895 Myc proto-oncogene protein Human genes 0.000 description 2
- 101710135898 Myc proto-oncogene protein Proteins 0.000 description 2
- 102100026715 Serine/threonine-protein kinase STK11 Human genes 0.000 description 2
- 101710150448 Transcriptional regulator Myc Proteins 0.000 description 2
- 102100039865 Ubiquitin carboxyl-terminal hydrolase 1 Human genes 0.000 description 2
- 102100031306 Ubiquitin carboxyl-terminal hydrolase 44 Human genes 0.000 description 2
- 102100039414 WD repeat-containing protein 48 Human genes 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 102100033065 cAMP-dependent protein kinase catalytic subunit beta Human genes 0.000 description 2
- 239000003560 cancer drug Substances 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 239000003054 catalyst Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 2
- 206010020718 hyperplasia Diseases 0.000 description 2
- 238000002991 immunohistochemical analysis Methods 0.000 description 2
- 238000003364 immunohistochemistry Methods 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 201000005243 lung squamous cell carcinoma Diseases 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 210000002569 neuron Anatomy 0.000 description 2
- 239000012188 paraffin wax Substances 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 208000000587 small cell lung carcinoma Diseases 0.000 description 2
- 230000037439 somatic mutation Effects 0.000 description 2
- 206010041823 squamous cell carcinoma Diseases 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 230000034512 ubiquitination Effects 0.000 description 2
- 238000010798 ubiquitination Methods 0.000 description 2
- 230000003827 upregulation Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 101150042041 wdr48 gene Proteins 0.000 description 2
- SJAPSPJRTQCDNO-UHFFFAOYSA-N 6-chloro-1,2-benzoxazol-3-one Chemical compound ClC1=CC=C2C(O)=NOC2=C1 SJAPSPJRTQCDNO-UHFFFAOYSA-N 0.000 description 1
- 102100022900 Actin, cytoplasmic 1 Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 102100034540 Adenomatous polyposis coli protein Human genes 0.000 description 1
- 108700028369 Alleles Proteins 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 102100029707 DnaJ homolog subfamily B member 4 Human genes 0.000 description 1
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 description 1
- 102100037964 E3 ubiquitin-protein ligase RING2 Human genes 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- 108010004889 Heat-Shock Proteins Proteins 0.000 description 1
- 102000002812 Heat-Shock Proteins Human genes 0.000 description 1
- 101000924577 Homo sapiens Adenomatous polyposis coli protein Proteins 0.000 description 1
- 101000866008 Homo sapiens DnaJ homolog subfamily B member 4 Proteins 0.000 description 1
- 101001095815 Homo sapiens E3 ubiquitin-protein ligase RING2 Proteins 0.000 description 1
- 101001057193 Homo sapiens Membrane-associated guanylate kinase, WW and PDZ domain-containing protein 1 Proteins 0.000 description 1
- 101000712958 Homo sapiens Ras association domain-containing protein 1 Proteins 0.000 description 1
- 101000628562 Homo sapiens Serine/threonine-protein kinase STK11 Proteins 0.000 description 1
- 101000651893 Homo sapiens Slit homolog 3 protein Proteins 0.000 description 1
- 101000702545 Homo sapiens Transcription activator BRG1 Proteins 0.000 description 1
- 101000740048 Homo sapiens Ubiquitin carboxyl-terminal hydrolase BAP1 Proteins 0.000 description 1
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- 101000740049 Latilactobacillus curvatus Bioactive peptide 1 Proteins 0.000 description 1
- 239000012097 Lipofectamine 2000 Substances 0.000 description 1
- 102000007530 Neurofibromin 1 Human genes 0.000 description 1
- 108010085793 Neurofibromin 1 Proteins 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- -1 P16 Proteins 0.000 description 1
- 108010011536 PTEN Phosphohydrolase Proteins 0.000 description 1
- 102000014160 PTEN Phosphohydrolase Human genes 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102000004245 Proteasome Endopeptidase Complex Human genes 0.000 description 1
- 108090000708 Proteasome Endopeptidase Complex Proteins 0.000 description 1
- 102000001253 Protein Kinase Human genes 0.000 description 1
- 229940123573 Protein synthesis inhibitor Drugs 0.000 description 1
- 239000013614 RNA sample Substances 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 238000010240 RT-PCR analysis Methods 0.000 description 1
- 102100033243 Ras association domain-containing protein 1 Human genes 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 108091058545 Secretory proteins Proteins 0.000 description 1
- 102000040739 Secretory proteins Human genes 0.000 description 1
- 101710181599 Serine/threonine-protein kinase STK11 Proteins 0.000 description 1
- 102100031027 Transcription activator BRG1 Human genes 0.000 description 1
- 102000015098 Tumor Suppressor Protein p53 Human genes 0.000 description 1
- 108010078814 Tumor Suppressor Protein p53 Proteins 0.000 description 1
- 108010083111 Ubiquitin-Protein Ligases Proteins 0.000 description 1
- 102000006275 Ubiquitin-Protein Ligases Human genes 0.000 description 1
- 108010066496 Ubiquitin-Specific Proteases Proteins 0.000 description 1
- 102000018390 Ubiquitin-Specific Proteases Human genes 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 201000008395 adenosquamous carcinoma Diseases 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000000540 analysis of variance Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 230000004009 axon guidance Effects 0.000 description 1
- HFACYLZERDEVSX-UHFFFAOYSA-N benzidine Chemical compound C1=CC(N)=CC=C1C1=CC=C(N)C=C1 HFACYLZERDEVSX-UHFFFAOYSA-N 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 229960004316 cisplatin Drugs 0.000 description 1
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
- 238000011260 co-administration Methods 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 239000003636 conditioned culture medium Substances 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 102000034356 gene-regulatory proteins Human genes 0.000 description 1
- 108091006104 gene-regulatory proteins Proteins 0.000 description 1
- 238000001114 immunoprecipitation Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000009940 knitting Methods 0.000 description 1
- 208000003849 large cell carcinoma Diseases 0.000 description 1
- 201000005249 lung adenocarcinoma Diseases 0.000 description 1
- 201000010893 malignant breast melanoma Diseases 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000002493 microarray Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- JTSLALYXYSRPGW-UHFFFAOYSA-N n-[5-(4-cyanophenyl)-1h-pyrrolo[2,3-b]pyridin-3-yl]pyridine-3-carboxamide Chemical compound C=1C=CN=CC=1C(=O)NC(C1=C2)=CNC1=NC=C2C1=CC=C(C#N)C=C1 JTSLALYXYSRPGW-UHFFFAOYSA-N 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000000414 obstructive effect Effects 0.000 description 1
- 238000006384 oligomerization reaction Methods 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 238000001259 photo etching Methods 0.000 description 1
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Substances [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 108060006633 protein kinase Proteins 0.000 description 1
- 239000000007 protein synthesis inhibitor Substances 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
- 238000000197 pyrolysis Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000013139 quantization Methods 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000012096 transfection reagent Substances 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 230000006663 ubiquitin-proteasome pathway Effects 0.000 description 1
Abstract
The application that the present invention relates to USP33 in tumour more particularly relates to USP33 as tumor prognosis and detects the application of marker and USP33 in tumour medicine exploitation.Data provided by the invention show that the decline of USP33 expression quantity is bad related to tumor prognosis in kinds of tumors patient；And USP33 albumen inhibits Nasopharyngeal neoplasms by regulating and controlling Slit-Robo signal path.Therefore, USP33 can be used as tumor prognosis detection marker, while USP33 can also be applied to the exploitation of tumour medicine, provide new method and scheme for the detection and treatment of tumour.
Description
Technical field
The present invention relates to pharmaceutical technology fields, and in particular to USP33 as tumor markers and its treats lung in preparation
Application in cancer drug.
Background technique
Lung cancer is the most common cancer, annual dead about 1,380,000 people in the whole world, while its 5 years survival rates are about 15%.Although
There are numerous treatment means at present, including operation, radiotherapy, chemotherapy and targeted therapy are even treated in combination between them, but it is imitated
Fruit is all barely satisfactory.Lung cancer can be divided into Small Cell Lung Cancer SCLC and non-small cell lung cancer NSCLC.Non-small cell lung cancer, including
Squamous cell carcinoma, gland cancer, undifferentiated large cell carcinoma, adenosquamous carcinoma and bronchial gland carcinoma；Small Cell Lung Cancer, by uniform small
Cell composition is typically in oat sample.NSCLC accounts for about 80% case.The treatment of current lung cancer is based primarily upon tumour shape
State and the Staging System based on moving TNM of being carried down by tumor lympha.Many tumor suppressor genes are advantageously, it has been found that include in lung cancer
TP53, P16, LKB1/STK11, NF1, RASSF1, APC, BRG1, PTEN and RB.Various tumor susceptibility genes also have been found recently,
However, the endogenetic mechanisms of invasion and the transfer of lung cancer is inhibited still to know little about it.
Recent report shows, neuron guide molecule is considered invasion to tumour and transfer process is related.Slit base
Because being initially found in drosophila, its product is a secretory protein family, serves as the effect of neural guide molecule.Nearest
Research shows that slit gene participates in infecting and shifting for different types of cancer cell.Slit albumen by with a single pass transmembrane
Albumen Roundabout's (Robo) is implemented in combination with its function.Regulate and control Slit-Robo signal transduction effect albumen first is that general
Plain specific protease 33 (USP33).An one of member as ubiquitin-specific protease family, USP33 is originally found
Protein binding as substrate is to VHL E3 ligase.In Slit signal path regulation middle line position joint axon guidance,
USP33 is considered required.
In addition, slit signal is in realizing the function that it inhibits breast cancer cell to migrate, USP33 is also assisted in wherein, this table
Bright USP33 may play a significant role in invasive cancer and transfer.However, how USP33 regulates and controls slit signal in lung cancer
The mechanism of conduction is not known.
Summary of the invention
It is prepared by the application that the technical problem to be solved by the invention is to provide USP33 in lung cancer, especially USP33
Application in detection and tumor.
To solve the above problems, the technical solution that first aspect present invention provides is: application of the USP33 in tumour,
USP33 detects the application of marker and USP33 in tumour medicine exploitation as tumor prognosis.
Preferably, USP33 as tumor prognosis detect marker, the tumour include lung cancer, breast cancer, melanoma and
Acute myelocytic leukemia.
Preferably, USP33 is lung cancer preparing the application in tumour medicine, the tumour.
Preferably, USP33 is as cancer target.
Preferably, USP33 inhibits Nasopharyngeal neoplasms by regulation slit-Robo signal path.
Second aspect of the present invention provides a kind of diagnostic kit, which contains detection USP33 protein substance, described
USP33 protein sequence is Seq No.1.
Third aspect present invention provides a kind of kit, which contains the substance of detection USP33 gene, described
USP33 gene order is Seq No.2.
Fourth aspect present invention provides a kind of purposes of diagnostic kit, for detecting tumour and tumor prognosis USP33 table
It reaches.
Fifth aspect present invention provides a kind of tumors pharmaceutical combination, which contains: USP33 gene and/or
Its albumen and medically acceptable pharmaceutic adjuvant that encode.
Preferably, the tumour is lung cancer.
During this investigation it turned out, inventor proves that Slit albumen inhibits the transition process of lung carcinoma cell that USP33 is needed to participate in.
USP33 adjusts the level of Robo receptor in lung carcinoma cell.This and USP33 have bright in the mode of action of neuron and non-cancerous cells
Aobvious difference (Yuasa-Kawada et al., 2009a, b).In addition, USP33 plays the part of tumor inhibitor in lung carcinoma cell
Role, because very strong correlativity is presented in the low expression of USP33 and the lower survival rate of the prognosis of patients with lung cancer.
USP33 expression is remarkably decreased in cancerous lung tissue.High USP33 expression is preferably related with prognosis.USP33 can be improved
Robo1 stability, and be that slit albumen inhibits necessary to tumor cell migration.These discoveries show that USP33 is lung cancer hair
The crucial participant of raw one, holds promise as a new lung cancer for prognosis marker.
USP33 provided by the invention can adjust the level of Robo receptor in lung carcinoma cell, and it is protein stabilized to improve Robo1
Property, and be that slit albumen inhibits necessary to tumor cell migration.Therefore USP33 can be used as treatment tumour, especially lung
The important component of cancer drug combines other anticancer drug co-administrations.In addition, USP33 is also used as lung cancer for prognosis detection mark
Will object provides new method and scheme for the detection and treatment of tumour, particularly lung cancer.
Detailed description of the invention
The invention will be further described with reference to the accompanying drawings and embodiments:
Fig. 1 shows the expression of mRNA in 25 groups of lung cancer and non-lung cancer tissue samples, and Y-axis is expression quantity, GAPDH base
Because doing reference, P < 0.001 * * *.
Fig. 2 immunohistochemical staining detects USP33 expression in lung cancer and check sample.Wherein, a and c is at normal group
Detect that the Strong positive signals of USP33 expression, b are the USP33 positive expression example in gland cancer in knitting, d is squamous cell carcinoma
USP33 positive expression example.
Fig. 3 lung cancer sample (N=25) and control normal tissue sample (CTRL) (N=25) USP33 protein expression situation
Quantitative analysis.
The calculation method of the USP33- immunohistochemistry score is as follows: USP33 immunohistochemistry score=(positive tumor cell
Percentage) × staining signals intensity.USP33 expressing quantity is substantially less than non-lung cancer tissue (* *, P < 0.01) in lung cancer.
Fig. 4 declines from the expression quantity for finding the USP33 in patients with lung cancer sample in the database of announcement through analysis；Wherein,
Fig. 4 A is USP33 genetic mutation and expression in TCGA database lung cancer sample.Light blue expression homozygosity lacks
It loses；Green indicates somatic mutation；Pink indicates expression up-regulation；Blue indicates that expression is lowered.The meter that these results are relied on
Count according to from cancer CBIO portal website ((http://www.oncomine.org).)
Fig. 4 B is USP33 amino acid mutation situation in TCGA database lung cancer sample.
Fig. 4 C is to analyze patients with lung cancer in five groups of databases using Oncomine (http://www.oncomine.org)
The expression of USP33 gene.In five groups of databases, compared with non-tumor sample, the expression quantity of USP33 occurs lung cancer sample
It is decreased obviously.LUAD: adenocarcinoma of lung；LuSC: squamous cell lung carcinoma, Ctrl: control.
Fig. 5 is USP33 changes in gene expression situation in different tissues sample.
Fig. 6, which is shown, shows that low expression USP33 gene means shorter life cycle in various cancers database；Specifically such as
Under
Fig. 6 A is TCGA lung cancer data library AgilentG4502A_07_3 data, n=91, USP33high in USP33low
Middle n=91；P=0.0436,
Fig. 6 B is TCGA lung cancer data library IlluminaHiSeq_RNASeqV2 data, n=106 in USP33low,
N=106 in USP33high；P=0.0267,
Fig. 6 C is that the data in GSE3141 lung cancer data library are by KM (http://kmplot.com/analysis/)
It is analyzed.N=83 in n=28 in USP33low, USP33high；P=0.031,
Fig. 6 D is that the data in GSE31210 lung cancer data library are by KM (http://kmplot.com/analysis/)
It is analyzed.N=113 in n=113 in USP33low, USP33high；P=0.034,
Fig. 6 E is n=61 in TCGA lung cancer data library USP33low, n=61 in USP33high；P=0.0188,
Fig. 6 F is lung cancer data Cooley n=562 in KM analysis USP33low, n=553 in USP33high；P=
0.034,
Fig. 6 G is n=145 in TCGA melanoma database USP33low, n=50 in USP33high；P=0.0453,
Fig. 6 H is TCGA acute myelocytic leukemia database, n=82 in n=77 in USP33low, USP33 high；P
=0.0282.
Fig. 7 A and 7B are respectively after H1299 cell is handled after transfecting siRNA or siUSP33 with control or slit albumen
Picture and analysis of accounts figure do not have significant change in third day and the 4th day control group and slit processing group cell quantity.
Fig. 8 is that USP33 passes through the adjusting slit inhibition lung carcinoma cell transfer of ubiquitination catalytic site；Wherein, Fig. 8 A is transfection
Compare the H1299cells wound healing assay picture of siRNA or siUSP33；Fig. 8 B and Fig. 8 E indicate cell migration distance.Figure
8C shows that Western test is proved to be siUSP33 rather than compareing siRNA can inhibit USP33 gene in H1299 cell
Expression.Fig. 8 D is wild type USP33 or mutation USP33 (C163A) be used to comfort control or slit control.Generate scratch
Afterwards, the cell at edge will be moved towards right side, and figure different time points after scratch acquire, and white dotted lines indicate to hurt for 0 hour
Mouth edge.Fig. 8 F is that Western verifies USP33 wild type and mutated gene (C163A) is expressed in H1299 cell.
Fig. 9 shows that USP33 and Robo1 interacts, and influences stability of the Robo1 in lung carcinoma cell.Wherein, scheme
The interaction of 9A USP33 and Robo1 in H1299 cell.Control IgG or Robo1 antibody is utilized in control and slit group
Carry out co-immunoprecipitation experiment.The albumen that immunoprecipitation obtains carries out Western blotting detection using USP33 antibody,
Fig. 9 B, which is shown, carries out transfection Robo-HA, Flag-ubiquitin, the H1299 of Ctrl siRNA or siUSP33
Deubiquitination experiment.After MG132 (20 μm of ol/L) handles 6h, co-immunoprecipitation experiment is carried out using HA antibody, later
Western blotting detection is carried out to obtained albumen,
Fig. 9 C shows that the H1299 cell of transfection Robo-HA, the USP33 and untransfected of wild type USP33 or mutation carry out
Deubiquitination situation.After MG132 (20 μm of ol/L) handles 6h, co-immunoprecipitation experiment is carried out using HA antibody, later
Western blotting detection is carried out to obtained albumen,
Fig. 9 D and Fig. 9 E are shown in transfection siRNAs USP33#1, USP33#2, compare Western blot after siRNAs
Expression of the USP33 and Robo1 in H1299 cell is detected,
Fig. 9 F show H1299 cell transfecting siUSP33 or control siRNA after with CHX (50 μ g/mL) processing it is different when
Between, and then detect the total amount in cell of Robo1.
Fig. 9 G, Fig. 9 H are handled after showing 1299 cell transfecting siUSP33 or control siRNA with MG132 (20 μm of ol/L)
6h (what is do not handled compares) carries out Western blot detection later.
Specific embodiment
The specific embodiment of the invention is described below in conjunction with attached drawing.
One, material
1. experimental animal or material source and processing
The sample that cancerous lung tissue and non-tumour lung tissue are collected after the diagnosis of The Fourth Military Medical University's Hospital Pathological Department room, owns
Sample standard deviation obtains patient's informed consent and in accordance with the guilding principle of associated mechanisms and country.As the following table 1 shows lung cancer sample distribution
Situation.
The tumor tissues sample of operation excision is used into rapidly liquid nitrogen cryopreservation, to extract RNA.
Cancerous lung tissue sample is fixed overnight through 10% neutral formalin solution, specimens paraffin embedding slices, according to standard scheme
H and E dyeing is carried out, immunohistochemical analysis is carried out.
1 lung cancer sample distribution of table
H1299 cell (comes from ATCC cell bank), and HEK293 cell (comes from ATCC cell bank), with 10% (v/v) tire ox
The DMEM culture medium culture of serum, 50mg/ml penicillin/streptomycin, temperature are 37 DEG C, gas concentration lwevel 5%.
2, drug and reagent:
2.1 drug
USP33 antibody is purchased from PROTEINTECH GROUP company, and Robo antibody is purchased from PROTEINTECH GROUP company,
Beta-actin antibody is purchased from PROTEINTECH GROUP company；Cycloheximide is purchased from Sigma company；MG132 is purchased from Sigma
Company.Trizol is purchased from Invitrogen company
2.2 reagent
Lipofectamine2000 transfection reagent is purchased from Invitrogen company；SiRNAs (the #1:5'- of USP33
UCUCGACAGUGGCUUAAUUAA-3',
#2:5'-GGAUUCAGUUGGUGAAAUUAC-3') and negative control siRNA (5 '-CGUACGCGGAAUA
CUUCGATT-3 ') it is synthesized by Shanghai JiMa pharmacy Technology Co., Ltd；SYBR green PCR mix is purchased from Thermo
Fisher Scientific company,
2.3 instrument
Ultrospec5300pro spectrophotometer is purchased from GE company；ScanScope XT scanner is purchased from U.S. Aperio
Company, real-time fluorescence quantitative PCR instrument are purchased from ABI company.
Packet data indicates plus or minus standard deviation from the mean.T examine (two samples) or ANOVA (more than two sample) into
The analysis of row statistical significance.Survival Analysis is drawn analysis curve and is analyzed using Prism software and log-rank.It is all
At least in triplicate, 95% confidence level of p < 0.05 is significant for experiment.
Two, experimental method and interpretation of result
1, in lung carcinoma cell and non-tumour lung tissue cell USP33 expression.
1.1 analyze USP33mRNA expression by quantitative RT-PCR method, compared with non-tumour lung tissue sample,
USP33 expression significantly reduces (P < 0.001) (Fig. 1).
RNA total in cancerous lung tissue is extracted using Trizol, using Ultrospec5300 spectrophotometer to the amount of RNA
It is measured with purity, removes genomic DNA from RNA sample with digestion DNA enzymatic I, draw using 5 microgram RNA as template oligomerization
Object (dT) and reverse transcriptase (M-MLV RT) synthesize the first chain of cDNA, are expanded using the StepOne real-time PCR of ABI
It is as follows to increase the primer:
USP33(5'-TGTGATGCTTAGGCAAGGAG-3',Seq No.3,
And 5'-GGCCCTCCACCATAAATAGA-3'), Seq No.4；
Robo1(5'-GCATCGCTGGAAGTAGCCATACT-3',Seq No.5
And 5'-CTAGAAATGGTGGGCTCAGGAT-3') Seq No.6；
GAPDH(5'-GGAGCGAGATCCCTCCAA AAT-3',Seq No.7
And 5'-GGCTGTTGTCATACTTCTCATGG-3'), Seq No.8.Amplification program:
95 DEG C, 15 seconds, 60 DEG C, 30 seconds, 72 DEG C, 30 seconds, totally 40 circulations, utilize 2-ΔΔCTMethod is calculated.
1.2 next, inventor by immunohistochemical staining method detect sample in USP33 protein expression situation (see Fig. 2
The specificity of anti-USP33).
Cancerous lung tissue sample is fixed through 10% formalin, 5 microns of thick tumor biopsies of paraffin embedding are immunized
Chemical analysis.It dewaxes first, rehydration heats 20 minutes in citric acid (pH value 6.0) micro-wave oven.After antigen retrieval, with 3% mistake
Hydrogen oxide is handled 10 minutes, closes endogenous peroxidase activity, then uses lowlenthal serum obstructive incubation in 30 minutes.
USP33 primary antibody is then incubated for biotin antibody to 4 DEG C of overnight incubations of slice, and room temperature 30 minutes.Finally, slice is through 3,3- diamino
The observation of base benzidine staining.Coloring slice is digitized through Scanscope XT.USP33 protein expression height is indicated with numerical value,
The average percent that positive tumor cell is determined after five random areas amplify × 200 is at least chosen in each slice.
USP33 intensity scores are as follows: 0, it is negative；1+, it is weak；2+, mild 3+, acutely.Positive tumour cell, the percentage of staining power
Then amplification generates USP33- immunohistochemical staining score.
In normal lung tissue, USP33 staining signals indication range is 73.33~300.00 (average values 198.18),
And the staining signals in those lung cancer samples change from 0.00 to 266.00 (average value 142.38).In lung cancer sample
USP33 expression is significantly lower than non-tumour lung tissue (P < 0.01, Fig. 2 and 3).
1.3 in order to investigate the variation that USP33 in patients with lung cancer is expressed be the inventor couple as caused by what reason
554 data are analyzed in cBioPortal cancer gene group database (http://cbioportal.org).Lung cancer
Survival and USP33 gene expression correlation are carried out using TCGA cancer databases and online KM-Plotter database
Analysis.
USP33mRNA is lowered in 9 cases of lung cancer, and 2 cases of lung cancer gene delections, in addition 9 body cell occur
The gene mutation (Fig. 4 A and 4B).It is worth noting that, three USP33 mutation (figures inside the catalyst structure domain of USP33
4B).Therefore, these cases of lung cancer about 4%, the expression for showing Homozygous deletions or somatic mutation or mRNA are lowered.
In addition, body cell USP33 missense mutation is sent out in the sample of adenocarcinoma of lung (4.3-6.2%) and squamous cell lung carcinoma (3.4%)
Existing (Fig. 5).Further the data set of USP33 express spectra in Oncomine database is analyzed (http: //
Www.oncomine.org/), which contains 5 groups of independence lung cancer sample mRNA gene table data.It is worth noting that, with right
Condition ratio in the same old way, in 5 groups of all data (Fig. 4 C), the USP33mRNA expression in lung cancer sample is significantly reduced.To sum up
Described, the downward of USP33 expression has high consistency in these lung cancer samples.
2, the expression and clinical effectiveness of USP33
In order to study the relationship between the expression of USP33 and clinic, inventor passes through disclosed micro-array chip number
It is analyzed according to the expression to USP33 and being associated between patient survival.Inventor is from cancer gene group map
Kaplan-Meier (KM) survivorship curve data and online KM- are generated in Cancer Genome Atlas (TCGA) data
Plotter database.For every group of data, according to the expression of USP33 gene, patients with lung cancer is divided into two groups.Kaplan-
Meier is used to analyze the life span difference of assessment USP33 high expression and USP33 low expression group.In the number of all four lung cancer
According to library, higher USP33 expression is with patient's longer life cycle (Fig. 6 A-6D).
Inventor further analyzes the relationship of USP33 expression Yu other kinds of cancer patient's life cycle.As a result it shows
Show, in other kinds of cancer, including breast cancer (Fig. 6 E and 6F), melanoma (Fig. 6 G) and acute myeloid leukemia
(AML, Fig. 6 H), shorter life cycle in a certain range with USP33 low expression level.These statistics indicate that,
In a variety of human cancers, USP33 plays the role of tumor suppressor gene.
3, USP33 participates in the inhibition lung carcinoma cell transfer that Slit signal is mediated
Inventor's previous studies show in different tumour cells (including lung cancer and breast cancer) slit protein expression
Amount is decreased obviously, and slit is played an important role in anti-curing cancers further develop.Existing result of study also turns out simultaneously,
In lung cancer, USP33 participates in the signal transduction of slit.In order to explore specific mechanism of action of the USP33 in lung carcinoma cell, invent
People has studied how it regulates and controls Slit signal path to inhibit the transfer of lung carcinoma cell.
3.1 recombination Slit2 productions and wound healing assay
Fusion is had to the slit2 gene transfected HEK 293 of c-myc label, Slit2 of the production with c-myc label is complete
Long albumen.Cell is cultivated in the DMEM culture medium of 5% fetal calf serum.Slit2 albumen is purified from culture supernatant.With what is do not transfected
HEK293 cell conditioned medium compares.
When carrying out wound healing assay, by H1299 cell kind on being coated with collagen photochemical etching grid slide,
These slides are placed in 35mm culture dish.Once cell aggregation is generated with the sterile careful cell scraping single layer of 10ul pipette tip
Wound area.Plate hole removal is rinsed with PBS and is detached from cell, is then cultivated in control group and culture medium containing Slit2.0 hour and
Wound forms 13-16 hours latter, and ten pictures are shot under inverted microscope.By measuring the distance of cell initial position forward
The wound that migration is formed carrys out the level of quantization cell migration.
Prove that Slit albumen can substantially reduce the cell migration of H1299 lung carcinoma cell by cell wound healing assay.
And this inhibiting effect USP33 by specific siRNA (siUSP33) knock out after disappear (Fig. 8 A-8C), it is such the result shows that
USP33 affects the migration of lung carcinoma cell by slit signal path.It should be noted simultaneously that in these experiments either
H1299 cell is handled with slit or siUSP33 acts on H1299 cell, does not influence the proliferation or cell week of H1299 cell
Phase (see Fig. 7).
In order to detect whether it is that the ubiquitin activity of going of USP33 has mediated this function, inventor uses USP33 mutant
It is tested.Inventor constructs USP33-C163A of a deubiquitination catalytic site point mutation, by wild type (Wt)
USP33 or mutant C163A-mutant USP33 (Mt) converts H1299 cell, later with the culture medium containing Slit or not
Control medium containing Slit is cultivated, and can be seen that in the presence of Slit from experimental result, the mutation of USP33
(C163A) it will increase the migration of H1299 cell.Thus inventor is it follows that USP33 albumen removes ubiquitin active structure domain pair
The migration of lung carcinoma cell is inhibited to play a significant role.
4, USP33 adjusts the stability of Robo1 albumen
Existing report display, in breast cancer cell, the effect of USP33 is to change the distribution of intracellular Robo1 albumen,
I.e. by Robo1Robo1 from being re-assigned to cell surface into the cell, the total level without will affect Robo1 albumen.So in lung
Whether USP33 has similar function in cancer cell.
Inventor has detected the interaction (Fig. 9 A) of USP33 and Robo1 with co-immunoprecipitation method, in H1299 cell
Middle missing USP33 gene, in the presence of MG-132 (inhibitor of Ubiquitin-proteasome systerm), the ubiquitination water of Robo1Robo1
It is flat to increase (Fig. 9 B).It is overexpressed wild type USP33 gene (rather than mutant C163A of catalytically inactive), the ubiquitin of Robo1
Change horizontal reduction (Fig. 9 C).It is surprising that lowering USP33 in H1299 cellular expression levels, Robo1 by siRNA method
Protein expression level also reduces, this (Fig.9D opposite with the result in breast cancer cell；Yuasa-Kawada et al.,
2009a).It is opposite, it is overexpressed the USP33 of wild type, rather than the USP33 of C163A mutation, the Robo1 table in H1299 cell
Increase (Fig. 9 E) up to amount.
In order to study the stability how USP33 maintains Robo1 albumen, inventor has carried out following experiment: with control
SiRNA (CTRL) or siUSP33 transfects H1299 cell, while with cycloheximide (CHX, protein synthesis inhibitor) to cell
Carry out processing in different time periods.Pass through Robo1 protein expression water in Western blotting (WB) analysis detection cell pyrolysis liquid
It is flat.6 hours after cycloheximide processing, the decline of Robo1 level；After processing 12 hours, compared with the siRNA of control, transfecting
Robo1 albumen is almost degraded in the H1299 cell of siUSP33, such the experimental results showed that under USP33 expression
Adjust the half-life period (Fig. 9 F) for shortening Robo1.The phenomenon that reducing Robo1 protein level by siUSP33, can be pressed down by proteasome
The experiment (Fig. 9 G) that preparation MG-132 is inhibited shows that Robo1 degradation mainly passes through Ubiquitin-proteasome in H1299 cell
What system was realized.These results indicate that Robo1 protein stability can be improved in USP33 in lung carcinoma cell, ubiquitin-egg is prevented
Degradation of the white enzyme body to it.
Three, interpretation of result
The evidence that the result of study of inventor provides shows that USP33 is a new member in lung cancer, adjusts slit letter
Number access.Inventors have demonstrated that the expression downward of USP33 is related to clinical effectiveness in lung cancer.USP33 is by adjusting slit letter
The activity suppression lung carcinoma cell migration of number access.In addition, USP33 improves Robo1 albumen by inhibiting protein degradation enzymatic activity
Stability.These results disclose USP33 and mediate the one not previously known effect of slit-robo signal in lung carcinoma cell.
USP33 participates in many cell processes, and the research of inventor proves that USP33 is a kind of lung cancer related gene for the first time,
And gene expression quantity in lung cancer tissue sample is to reduce.This downward USP33 expression is in the mostly micro- battle array of lung cancer
In column database, and confirmed by real-time quantitative RT-PCR and immunohistochemical analysis.The high expression of USP33 and lung cancer
Patient's prognosis is preferably related.Mankind's USP33 gene is located at No. 1 chromosome p31.1, in non-small cell lung cancer there are about 50% should
Region deletion allele.In this region, some tumor suppressor genes have been determined.For example, with DnaJ-like heat shock protein
White (HLJ1) is located in this region and has been identified as inhibiting the proliferation of lung carcinoma cell, Anchorage Independent growth, and tumour occurs,
Cell transfer and invasion.CAMP- deopendent protein kinase β (PRKACB) is lowered in non-small cell lung cancer (NSCLC) tissue,
And the up-regulation of PRKACB can prevent the development of non-small cell lung cancer.In our current research, the result of study of inventor shows
USP33 is the key that participant occurs for lung cancer, can be a potential prognosis biomarker as prediction lung cancer development.
It has recently been demonstrated that different genes are expressed in ubiquitin protein family change and different types of cancer phase
It closes, including lung cancer etc..For example, USP44 is lowered in human lung adenocarcinoma.Patient USP44 expression of enzymes level is lower to be shown always to survive
Phase will substantially reduce.In lung cancer cell line it has been found that this goes in ubiquitin (BAP1) homozygote to reset and lack.USP1/UAF1 can be with
As a kind of oncogene, because the inhibitor and cis-platinum of USP1/UAF1 are inhibiting non-small cell lung cancer cisplatin-resistant cell to increase
Grow middle performance synergistic effect.Present patent application research shows that: USP33 may be a kind of new tumor suppressor gene in lung cancer.
In Human Lung Cancer, the low expression of Slit2 albumen and the state of an illness prognosis of advanced lung cancer and patient are bad related.
The mouse of Robo1 defect shows bronchiolar epithelium hyperplasia and focal hyperplasia, the i.e. relevant pathological characteristic of the early stage of lung cancer.It is sending out
In the research of bright people, Slit2 albumen significantly inhibits the migration of lung carcinoma cell, and slit-Robo signal path is supported to have suppression to lung cancer
Production is used.
The research of inventor has shown that slit inhibits effect of the breast cancer cell migration dependent on USP33, in lung carcinoma cell
In, USP33 and Robo1 interact, and are necessary to the migration of slit inhibition lung carcinoma cell.The modification that ubiquitin mediates is made
Play a key effect (Ciechanover and Schwartz, 1994) in the stability of regulatory protein.However, USP33
It interacts with Robo1, is that the Robo1 that slit is mediated is redistributed to required for plasma membrane, but does not influence total Robo1 albumen
It is horizontal.
Show USP33 by inhibiting ubiquitin-proteasome pathway thin in lung cancer to stablize Robo1 albumen in this research
Expression in born of the same parents.USP33 regulates and controls conduction of the slit signal in lung carcinoma cell, this needs the enzymatic activity of USP33, because
After the mutation of USP33 catalytic site (C163A), slit no longer inhibits lung carcinoma cell to migrate.It is interesting that in cBioPortal lung cancer
4 catalyst structure domain missense mutation in USP33 are had determined that in sample genomic database.Synthesis is special to the invention of breast cancer
Benefit application research can be shown that in different cancer cells, USP33 may pass through different mechanism regulating slit signal transductions.
The basic principles, main features and advantages of the present invention have been shown and described above.The technology of the industry
Personnel only illustrate the present invention it should be appreciated that the present invention is not limited by examples detailed above described in examples detailed above and specification
Principle, various changes and improvements may be made to the invention without departing from the spirit and scope of the present invention, these variation and
Improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention is by appended claims and its is equal
Object defines.
Claims (3)
- Application of the 1.USP33 in preparation detection tumor prognosis development drug, the tumour is lung cancer, melanoma and urgency Acute myeloid leukemia, the prognosis refer to the life cycle of patient.
- 2. application according to claim 1, it is characterised in that: the USP33 is as tumor suppressor gene.
- 3. application according to claim 2, it is characterised in that: in lung cancer, the USP33 passes through regulation slit-Robo Signal path is as tumor suppressor gene.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410242697.3A CN105214077B (en) | 2014-06-03 | 2014-06-03 | Application of the USP33 in tumour |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410242697.3A CN105214077B (en) | 2014-06-03 | 2014-06-03 | Application of the USP33 in tumour |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105214077A CN105214077A (en) | 2016-01-06 |
| CN105214077B true CN105214077B (en) | 2019-02-05 |
Family
ID=54983587
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410242697.3A Active CN105214077B (en) | 2014-06-03 | 2014-06-03 | Application of the USP33 in tumour |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105214077B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017192965A2 (en) * | 2016-05-05 | 2017-11-09 | Integrated Diagnostics, Inc. | Compositions, methods and kits for diagnosis of lung cancer |
| CN111748531B (en) * | 2019-03-29 | 2022-04-26 | 中国科学院北京基因组研究所(国家生物信息中心) | Application of USP33 as medication target in preparation of medicines |
| CN114231635B (en) * | 2021-12-31 | 2023-03-31 | 博尔诚（北京）科技有限公司 | Marker and probe composition for lung cancer screening and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101283106A (en) * | 2005-07-27 | 2008-10-08 | 肿瘤疗法科学股份有限公司 | Method of diagnosing small cell lung cancer |
| CN101495498A (en) * | 2005-02-07 | 2009-07-29 | 基因信息公司 | Mild osteoarthritis biomarkers and uses thereof |
| WO2012031008A2 (en) * | 2010-08-31 | 2012-03-08 | The General Hospital Corporation | Cancer-related biological materials in microvesicles |
| WO2013070521A1 (en) * | 2011-11-08 | 2013-05-16 | Genomic Health, Inc. | Method of predicting breast cancer prognosis |
| CN103890193A (en) * | 2011-08-29 | 2014-06-25 | 心脏Dx公司 | Methods and compositions for determining smoking status |
-
2014
- 2014-06-03 CN CN201410242697.3A patent/CN105214077B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101495498A (en) * | 2005-02-07 | 2009-07-29 | 基因信息公司 | Mild osteoarthritis biomarkers and uses thereof |
| CN101283106A (en) * | 2005-07-27 | 2008-10-08 | 肿瘤疗法科学股份有限公司 | Method of diagnosing small cell lung cancer |
| WO2012031008A2 (en) * | 2010-08-31 | 2012-03-08 | The General Hospital Corporation | Cancer-related biological materials in microvesicles |
| CN103890193A (en) * | 2011-08-29 | 2014-06-25 | 心脏Dx公司 | Methods and compositions for determining smoking status |
| WO2013070521A1 (en) * | 2011-11-08 | 2013-05-16 | Genomic Health, Inc. | Method of predicting breast cancer prognosis |
Non-Patent Citations (2)
| Title |
|---|
| Deubiquitinating enzyme USP33/VDU1 is required for Slit signaling in inhibiting breast cancer cell migration;Junichi Yuasa-Kawada等;《Proceedings of the National Academy of Sciences of the United States of America》;20090825;第106卷(第34期);全文 |
| 登录号：NM_201624;Curcio-Morelli,C.等;《Genbank》;20040210;序列信息 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105214077A (en) | 2016-01-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1890381B (en) | Method of diagnosing breast cancer | |
| Murali et al. | Tumours associated with BAP1 mutations | |
| JP2007506425A (en) | Methods for diagnosing hepatocellular carcinoma | |
| JP4851451B2 (en) | Breast cancer-related gene ZNFN3A1 | |
| EP2681330B1 (en) | Use of the olfactomedin-4 protein (olfm4) in colorectal cancer diagnosis | |
| JP2006500946A (en) | Diagnostic method of testicular seminoma | |
| Franks et al. | Unique roles of Akt1 and Akt2 in IGF-IR mediated lung tumorigenesis | |
| Li et al. | Phospholipase Cγ1 (PLCG1) overexpression is associated with tumor growth and poor survival in IDH wild-type lower-grade gliomas in adult patients | |
| Kyjacova et al. | IER2-induced senescence drives melanoma invasion through osteopontin | |
| TWI567391B (en) | Biomarker for identifying subgroup of early-stage lung adenocarcinoma patients | |
| CN109735620A (en) | A kind of application of molecular target in esophageal squamous cell carcinoma prognosis evaluation and treatment | |
| CN105214077B (en) | Application of the USP33 in tumour | |
| JP4614952B2 (en) | Hypoxia-inducible protein 2 (HIG2) as a potential new therapeutic target for renal cell carcinoma (RCC) | |
| CN101273131A (en) | Pancreatic cancer related gene CST6 and GABRP | |
| US11510911B2 (en) | Method for prediction of susceptibility to sorafenib treatment by using SULF2 gene, and composition for treatment of cancer comprising SULF2 inhibitor | |
| Schettino et al. | The potential role of pharmacogenomic and genomic in the adjuvant treatment of early stage non small cell lung cancer | |
| KR101793175B1 (en) | Method for Prediction of Susceptibility to Sorafenib Using SULF2 Gene and Composition for Treating Cancer Containing SULF2 Inhibitor | |
| US20210308213A1 (en) | Method for diagnosing and treating cancer using naïve state stem cell specific genes | |
| Zhang et al. | Hsa_circ_0094606 promotes malignant progression of prostate cancer by inducing M2 polarization of macrophages through PRMT1-mediated arginine methylation of ILF3 | |
| CN116121393B (en) | Marker for head and neck squamous cell carcinoma and application thereof | |
| EP3784240B1 (en) | Wnt6 as glioblastoma oncogenic biomarker, and uses of inhibitors thereof for treating wnt6-overexpressing glioblastoma | |
| RU2813996C2 (en) | Fusion protein of dctn1 protein with ret protein | |
| Dong et al. | The ‘Other’subfamily of HECT E3 ubiquitin ligases evaluate the tumour immune microenvironment and prognosis in patients with hepatocellular carcinoma | |
| Speransky | Identifying Novel Biomarkers of Metastatic Potential in Prostate and Breast Cancer | |
| CN106554962A (en) | Prevention, diagnosis and the treatment of the cancer of overexpression GPR160 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20181122 Address after: 315000 Gulin Section 2, Yinxian Avenue, Gulin Town, Haishu District, Ningbo City, Zhejiang Province Applicant after: ZHEJIANG ASCLEPLUS BIOTECHNOLOGY Co.,Ltd. Address before: 215028 128 Xinghu Street, Suzhou City, Jiangsu Province Applicant before: SUZHOU SHUNSHENGQIAO BIOTECHNOLOGY Co.,Ltd. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: Application of USP33 in tumors Effective date of registration: 20200413 Granted publication date: 20190205 Pledgee: Gulin sub branch of Ningbo Yinzhou Rural Commercial Bank Co.,Ltd. Pledgor: ZHEJIANG ASCLEPLUS BIOTECHNOLOGY Co.,Ltd. Registration number: Y2020330000160 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20220128 Granted publication date: 20190205 Pledgee: Gulin sub branch of Ningbo Yinzhou Rural Commercial Bank Co.,Ltd. Pledgor: ZHEJIANG ASCLEPLUS BIOTECHNOLOGY Co.,Ltd. Registration number: Y2020330000160 |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20220222 Address after: No. 129-8, Chuanjian South 1st Road, Jinniu District, Chengdu, Sichuan 610081 Patentee after: Sichuan asikeli Biotechnology Co.,Ltd. Address before: 215123 a2-425, bio nano Industrial Park, 218 Xinghu street, Suzhou Industrial Park, Jiangsu Province Patentee before: ASCLEPIUS (SUZHOU) TECHNOLOGY COMPANY GROUP Co.,Ltd. Effective date of registration: 20220222 Address after: 215123 a2-425, bio nano Industrial Park, 218 Xinghu street, Suzhou Industrial Park, Jiangsu Province Patentee after: ASCLEPIUS (SUZHOU) TECHNOLOGY COMPANY GROUP Co.,Ltd. Address before: 315000 Gulin Section 2, Yinxian Avenue, Gulin Town, Haishu District, Ningbo City, Zhejiang Province Patentee before: ZHEJIANG ASCLEPLUS BIOTECHNOLOGY Co.,Ltd. |