CN110904108B - 微小rna及其在制备抗肿瘤药物中的应用 - Google Patents
微小rna及其在制备抗肿瘤药物中的应用 Download PDFInfo
- Publication number
- CN110904108B CN110904108B CN201911293658.5A CN201911293658A CN110904108B CN 110904108 B CN110904108 B CN 110904108B CN 201911293658 A CN201911293658 A CN 201911293658A CN 110904108 B CN110904108 B CN 110904108B
- Authority
- CN
- China
- Prior art keywords
- artificial sequence
- rna
- cancer
- tumor
- microrna
- 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
- 239000002246 antineoplastic agent Substances 0.000 title claims description 14
- 229940041181 antineoplastic drug Drugs 0.000 title claims description 12
- 238000002360 preparation method Methods 0.000 title claims description 5
- 108700011259 MicroRNAs Proteins 0.000 title description 29
- 230000000295 complement effect Effects 0.000 claims description 20
- 206010028980 Neoplasm Diseases 0.000 claims description 15
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 12
- 206010017758 gastric cancer Diseases 0.000 claims description 12
- 201000011549 stomach cancer Diseases 0.000 claims description 12
- 239000002773 nucleotide Substances 0.000 claims description 10
- 125000003729 nucleotide group Chemical group 0.000 claims description 10
- 210000004881 tumor cell Anatomy 0.000 abstract description 13
- 230000012010 growth Effects 0.000 abstract description 11
- 230000000259 anti-tumor effect Effects 0.000 abstract description 9
- 230000035755 proliferation Effects 0.000 abstract description 5
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 76
- 210000004027 cell Anatomy 0.000 description 41
- 239000002679 microRNA Substances 0.000 description 26
- 108091070501 miRNA Proteins 0.000 description 16
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 10
- 201000005202 lung cancer Diseases 0.000 description 10
- 208000020816 lung neoplasm Diseases 0.000 description 10
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 8
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 8
- 201000007270 liver cancer Diseases 0.000 description 8
- 208000005016 Intestinal Neoplasms Diseases 0.000 description 7
- 201000002313 intestinal cancer Diseases 0.000 description 7
- 208000014018 liver neoplasm Diseases 0.000 description 7
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 6
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 6
- 229940079593 drug Drugs 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 230000014509 gene expression Effects 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 238000012986 modification Methods 0.000 description 6
- 230000004048 modification Effects 0.000 description 6
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 description 5
- 201000011510 cancer Diseases 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 5
- 208000003950 B-cell lymphoma Diseases 0.000 description 4
- 125000000217 alkyl group Chemical group 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 4
- 229940104302 cytosine Drugs 0.000 description 4
- 239000003112 inhibitor Substances 0.000 description 4
- -1 miR-27a Proteins 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 150000003290 ribose derivatives Chemical class 0.000 description 4
- 229930024421 Adenine Natural products 0.000 description 3
- 208000010839 B-cell chronic lymphocytic leukemia Diseases 0.000 description 3
- 206010009944 Colon cancer Diseases 0.000 description 3
- 208000031422 Lymphocytic Chronic B-Cell Leukemia Diseases 0.000 description 3
- 208000034578 Multiple myelomas Diseases 0.000 description 3
- 206010035226 Plasma cell myeloma Diseases 0.000 description 3
- 239000012980 RPMI-1640 medium Substances 0.000 description 3
- 229960000643 adenine Drugs 0.000 description 3
- 208000032852 chronic lymphocytic leukemia Diseases 0.000 description 3
- 230000034994 death Effects 0.000 description 3
- 208000005017 glioblastoma Diseases 0.000 description 3
- 201000010536 head and neck cancer Diseases 0.000 description 3
- 208000014829 head and neck neoplasm Diseases 0.000 description 3
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- 239000002502 liposome Substances 0.000 description 3
- 201000001441 melanoma Diseases 0.000 description 3
- 108091029119 miR-34a stem-loop Proteins 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 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
- FZWGECJQACGGTI-UHFFFAOYSA-N 2-amino-7-methyl-1,7-dihydro-6H-purin-6-one Chemical compound NC1=NC(O)=C2N(C)C=NC2=N1 FZWGECJQACGGTI-UHFFFAOYSA-N 0.000 description 2
- RYVNIFSIEDRLSJ-UHFFFAOYSA-N 5-(hydroxymethyl)cytosine Chemical compound NC=1NC(=O)N=CC=1CO RYVNIFSIEDRLSJ-UHFFFAOYSA-N 0.000 description 2
- HCGHYQLFMPXSDU-UHFFFAOYSA-N 7-methyladenine Chemical compound C1=NC(N)=C2N(C)C=NC2=N1 HCGHYQLFMPXSDU-UHFFFAOYSA-N 0.000 description 2
- MSSXOMSJDRHRMC-UHFFFAOYSA-N 9H-purine-2,6-diamine Chemical compound NC1=NC(N)=C2NC=NC2=N1 MSSXOMSJDRHRMC-UHFFFAOYSA-N 0.000 description 2
- LRFVTYWOQMYALW-UHFFFAOYSA-N 9H-xanthine Chemical compound O=C1NC(=O)NC2=C1NC=N2 LRFVTYWOQMYALW-UHFFFAOYSA-N 0.000 description 2
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 description 2
- 208000014697 Acute lymphocytic leukaemia Diseases 0.000 description 2
- 208000031261 Acute myeloid leukaemia Diseases 0.000 description 2
- 206010073478 Anaplastic large-cell lymphoma Diseases 0.000 description 2
- 201000003076 Angiosarcoma Diseases 0.000 description 2
- 206010003571 Astrocytoma Diseases 0.000 description 2
- 208000032791 BCR-ABL1 positive chronic myelogenous leukemia Diseases 0.000 description 2
- 206010005003 Bladder cancer Diseases 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 2
- 208000026310 Breast neoplasm Diseases 0.000 description 2
- 206010008342 Cervix carcinoma Diseases 0.000 description 2
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 2
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 2
- 208000010833 Chronic myeloid leukaemia Diseases 0.000 description 2
- 206010014733 Endometrial cancer Diseases 0.000 description 2
- 206010014759 Endometrial neoplasm Diseases 0.000 description 2
- 208000000461 Esophageal Neoplasms Diseases 0.000 description 2
- 208000032612 Glial tumor Diseases 0.000 description 2
- 206010018338 Glioma Diseases 0.000 description 2
- 208000001258 Hemangiosarcoma Diseases 0.000 description 2
- 208000017604 Hodgkin disease Diseases 0.000 description 2
- 208000021519 Hodgkin lymphoma Diseases 0.000 description 2
- 208000010747 Hodgkins lymphoma Diseases 0.000 description 2
- 208000007766 Kaposi sarcoma Diseases 0.000 description 2
- 208000032004 Large-Cell Anaplastic Lymphoma Diseases 0.000 description 2
- 206010023856 Laryngeal squamous cell carcinoma Diseases 0.000 description 2
- 208000018142 Leiomyosarcoma Diseases 0.000 description 2
- 208000025205 Mantle-Cell Lymphoma Diseases 0.000 description 2
- 208000000172 Medulloblastoma Diseases 0.000 description 2
- 206010027406 Mesothelioma Diseases 0.000 description 2
- 206010027476 Metastases Diseases 0.000 description 2
- 201000003793 Myelodysplastic syndrome Diseases 0.000 description 2
- 208000033761 Myelogenous Chronic BCR-ABL Positive Leukemia Diseases 0.000 description 2
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 description 2
- OVRNDRQMDRJTHS-CBQIKETKSA-N N-Acetyl-D-Galactosamine Chemical compound CC(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-CBQIKETKSA-N 0.000 description 2
- MBLBDJOUHNCFQT-UHFFFAOYSA-N N-acetyl-D-galactosamine Natural products CC(=O)NC(C=O)C(O)C(O)C(O)CO MBLBDJOUHNCFQT-UHFFFAOYSA-N 0.000 description 2
- 201000004404 Neurofibroma Diseases 0.000 description 2
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 description 2
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 2
- 206010033128 Ovarian cancer Diseases 0.000 description 2
- 206010061535 Ovarian neoplasm Diseases 0.000 description 2
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 2
- 208000006664 Precursor Cell Lymphoblastic Leukemia-Lymphoma Diseases 0.000 description 2
- 206010060862 Prostate cancer Diseases 0.000 description 2
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 2
- 208000015634 Rectal Neoplasms Diseases 0.000 description 2
- 108091028664 Ribonucleotide Proteins 0.000 description 2
- 206010041067 Small cell lung cancer Diseases 0.000 description 2
- 208000000102 Squamous Cell Carcinoma of Head and Neck Diseases 0.000 description 2
- 208000024313 Testicular Neoplasms Diseases 0.000 description 2
- 208000024770 Thyroid neoplasm Diseases 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 2
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 2
- DZBUGLKDJFMEHC-UHFFFAOYSA-N acridine Chemical compound C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 2
- 125000000304 alkynyl group Chemical group 0.000 description 2
- 125000004103 aminoalkyl group Chemical group 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 201000010881 cervical cancer Diseases 0.000 description 2
- 238000002512 chemotherapy Methods 0.000 description 2
- 208000029742 colonic neoplasm Diseases 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 201000004101 esophageal cancer Diseases 0.000 description 2
- 208000015419 gastrin-producing neuroendocrine tumor Diseases 0.000 description 2
- 201000000052 gastrinoma Diseases 0.000 description 2
- 125000001475 halogen functional group Chemical group 0.000 description 2
- 208000006359 hepatoblastoma Diseases 0.000 description 2
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 2
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 208000032839 leukemia Diseases 0.000 description 2
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 2
- 206010027191 meningioma Diseases 0.000 description 2
- 230000009401 metastasis Effects 0.000 description 2
- 108091062762 miR-21 stem-loop Proteins 0.000 description 2
- 108091041631 miR-21-1 stem-loop Proteins 0.000 description 2
- 108091044442 miR-21-2 stem-loop Proteins 0.000 description 2
- 230000003278 mimic effect Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 210000004877 mucosa Anatomy 0.000 description 2
- 208000025113 myeloid leukemia Diseases 0.000 description 2
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 2
- 201000008968 osteosarcoma Diseases 0.000 description 2
- 201000002528 pancreatic cancer Diseases 0.000 description 2
- 208000008443 pancreatic carcinoma Diseases 0.000 description 2
- 201000010198 papillary carcinoma Diseases 0.000 description 2
- 208000028591 pheochromocytoma Diseases 0.000 description 2
- 206010038038 rectal cancer Diseases 0.000 description 2
- 201000001275 rectum cancer Diseases 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 201000009410 rhabdomyosarcoma Diseases 0.000 description 2
- 239000002336 ribonucleotide Substances 0.000 description 2
- 125000002652 ribonucleotide group Chemical group 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 208000000649 small cell carcinoma Diseases 0.000 description 2
- 208000000587 small cell lung carcinoma Diseases 0.000 description 2
- 125000001424 substituent group Chemical group 0.000 description 2
- 201000003120 testicular cancer Diseases 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 description 2
- 201000002510 thyroid cancer Diseases 0.000 description 2
- 208000013077 thyroid gland carcinoma Diseases 0.000 description 2
- 230000014616 translation Effects 0.000 description 2
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 2
- 229940035893 uracil Drugs 0.000 description 2
- 201000005112 urinary bladder cancer Diseases 0.000 description 2
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- WKMPTBDYDNUJLF-UHFFFAOYSA-N 2-fluoroadenine Chemical compound NC1=NC(F)=NC2=C1N=CN2 WKMPTBDYDNUJLF-UHFFFAOYSA-N 0.000 description 1
- 125000003903 2-propenyl group Chemical group [H]C([*])([H])C([H])=C([H])[H] 0.000 description 1
- OVONXEQGWXGFJD-UHFFFAOYSA-N 4-sulfanylidene-1h-pyrimidin-2-one Chemical compound SC=1C=CNC(=O)N=1 OVONXEQGWXGFJD-UHFFFAOYSA-N 0.000 description 1
- LRSASMSXMSNRBT-UHFFFAOYSA-N 5-methylcytosine Chemical compound CC1=CNC(=O)N=C1N LRSASMSXMSNRBT-UHFFFAOYSA-N 0.000 description 1
- MIHCRNZMESVPJI-UHFFFAOYSA-N 5-sulfanyl-1h-pyrimidine-2,4-dione Chemical compound SC1=CNC(=O)NC1=O MIHCRNZMESVPJI-UHFFFAOYSA-N 0.000 description 1
- HRYKDUPGBWLLHO-UHFFFAOYSA-N 8-azaadenine Chemical compound NC1=NC=NC2=NNN=C12 HRYKDUPGBWLLHO-UHFFFAOYSA-N 0.000 description 1
- LPXQRXLUHJKZIE-UHFFFAOYSA-N 8-azaguanine Chemical compound NC1=NC(O)=C2NN=NC2=N1 LPXQRXLUHJKZIE-UHFFFAOYSA-N 0.000 description 1
- 229960005508 8-azaguanine Drugs 0.000 description 1
- FJNCXZZQNBKEJT-UHFFFAOYSA-N 8beta-hydroxymarrubiin Natural products O1C(=O)C2(C)CCCC3(C)C2C1CC(C)(O)C3(O)CCC=1C=COC=1 FJNCXZZQNBKEJT-UHFFFAOYSA-N 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- 229930183010 Amphotericin Natural products 0.000 description 1
- QGGFZZLFKABGNL-UHFFFAOYSA-N Amphotericin A Natural products OC1C(N)C(O)C(C)OC1OC1C=CC=CC=CC=CCCC=CC=CC(C)C(O)C(C)C(C)OC(=O)CC(O)CC(O)CCC(O)C(O)CC(O)CC(O)(CC(O)C2C(O)=O)OC2C1 QGGFZZLFKABGNL-UHFFFAOYSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 108020004394 Complementary RNA Proteins 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 1
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108091033773 MiR-155 Proteins 0.000 description 1
- 108091030146 MiRBase Proteins 0.000 description 1
- 108091027766 Mir-143 Proteins 0.000 description 1
- 108091028684 Mir-145 Proteins 0.000 description 1
- 241000204031 Mycoplasma Species 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 1
- 230000007022 RNA scission Effects 0.000 description 1
- 208000006265 Renal cell carcinoma Diseases 0.000 description 1
- 206010061934 Salivary gland cancer Diseases 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- 206010057644 Testis cancer Diseases 0.000 description 1
- 108091036066 Three prime untranslated region Proteins 0.000 description 1
- 208000033781 Thyroid carcinoma Diseases 0.000 description 1
- 102000044209 Tumor Suppressor Genes Human genes 0.000 description 1
- 108700025716 Tumor Suppressor Genes Proteins 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 125000003342 alkenyl group Chemical group 0.000 description 1
- 125000002877 alkyl aryl group Chemical group 0.000 description 1
- 229940009444 amphotericin Drugs 0.000 description 1
- APKFDSVGJQXUKY-INPOYWNPSA-N amphotericin B Chemical compound O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1/C=C/C=C/C=C/C=C/C=C/C=C/C=C/[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 APKFDSVGJQXUKY-INPOYWNPSA-N 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 229940034982 antineoplastic agent Drugs 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 125000003710 aryl alkyl group Chemical group 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000033228 biological regulation Effects 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
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003184 complementary RNA Substances 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- CHNUOJQWGUIOLD-NFZZJPOKSA-N epalrestat Chemical compound C=1C=CC=CC=1\C=C(/C)\C=C1/SC(=S)N(CC(O)=O)C1=O CHNUOJQWGUIOLD-NFZZJPOKSA-N 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 229910052731 fluorine Inorganic materials 0.000 description 1
- 239000011737 fluorine Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 125000000592 heterocycloalkyl group Chemical group 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000000138 intercalating agent Substances 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 125000005647 linker group Chemical group 0.000 description 1
- 125000001921 locked nucleotide group Chemical group 0.000 description 1
- 210000003563 lymphoid tissue Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- 108091037473 miR-103 stem-loop Proteins 0.000 description 1
- 108091057645 miR-15 stem-loop Proteins 0.000 description 1
- 108091027943 miR-16 stem-loop Proteins 0.000 description 1
- 108091007431 miR-29 Proteins 0.000 description 1
- 208000025189 neoplasm of testis Diseases 0.000 description 1
- 208000007538 neurilemmoma Diseases 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical group 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 238000009520 phase I clinical trial Methods 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 201000003804 salivary gland carcinoma Diseases 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 206010039667 schwannoma Diseases 0.000 description 1
- 125000003808 silyl group Chemical group [H][Si]([H])([H])[*] 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 150000003573 thiols Chemical class 0.000 description 1
- 229940113082 thymine Drugs 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000004565 tumor cell growth Effects 0.000 description 1
- 210000001635 urinary tract Anatomy 0.000 description 1
- 229940075420 xanthine Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
- C12N15/1135—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against oncogenes or tumor suppressor genes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/14—Type of nucleic acid interfering N.A.
- C12N2310/141—MicroRNAs, miRNAs
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Public Health (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oncology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Hematology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明涉及生物技术领域，特别涉及RNA或其应用。本发明提供的RNA分子长度为22个碱基；其5’端第1位～第6位碱基序列为“UGGCAG”，或者第17位～第22位碱基为“UGCCCU”；采用该RNA分子能够显著抑制肿瘤细胞生长和/或增殖，从而起到抗肿瘤的作用。
Description
技术领域
本发明涉及生物技术领域，特别涉及微小RNA及其在制备抗肿瘤药物中的应用。
背景技术
恶性肿瘤严重威胁人类健康和生命，已经成为我国城乡居民的首要死因。其中，发病率和死亡率最高的是肺癌，其次是肝癌、结直肠癌、胃癌。相关统计学资料显示，2015年全国新发恶性肿瘤392.9万例，发病率为285.83/10万；由于肿瘤疾病造成的死亡病例达到233.8万例，死亡率为170.05/10万。根据国际癌症研究署预测，如果不采取有效措施，到2020年我国癌症发病例数将达到400万，死亡人数将达到300万，肿瘤的发生对人们的生活质量和生存质量均构成了严重威胁，给患者家庭和社会造成了沉重的负担。目前治疗恶性肿瘤的主要方法有手术切除、药物化疗、放射治疗等，而由于大多数恶性肿瘤患者在发现肿瘤时已有转移，故药物化疗是必不可少的治疗方式之一。
微小RNA(microRNA)是近年来发现于真核细胞中的一类长约22个核苷酸的内源性非编码小分子RNA，其5′-端第2-9个碱基(seed region)可通过与靶基因的3′-UTR结合，在翻译水平抑制蛋白合成，从而在基因表达中发挥重要的调节作用。迄今为止，人类基因组中已明确的微小RNA约有2588个(miRBase)，调控着至少30％基因的表达，而每一个微小RNA可能参与调控100～200个靶基因的翻译。微小RNA由于其广泛的调控作用参与了细胞的生长、分化、增殖和凋亡等生命过程，影响着几乎所有的信号通路，并参与各种生理病理过程，特别在肿瘤的发生和发展中发挥了极为重要的作用。
大量研究结果显示微小RNA在肿瘤中呈现异常表达，其中有的微小RNA表达升高而有的降低，分别通过抑制抑癌基因表达或上调癌基因表达，发挥着促癌或抑癌的作用。2002年，Clain等发现两个微小RNA基因miR-15和miR-16在慢性淋巴细胞白血病患者中常发生缺失，首次揭示了微小RNA与肿瘤的密切关系。之后，越来越多的微小RNA被发现在肿瘤中异常表达，如低表达的miR-34a、miR-143、miR-145和高表达的miR-21、miR-27a、miR-155等。业已证实，通过在肿瘤细胞中转入低表达微小RNA的模拟物(mimic和agomir等)，或者转入高表达微小RNA的抑制剂(inhibitor和antagomir等)，可以有效抑制肿瘤细胞增殖、侵袭和转移，从而发挥抗肿瘤效果。
在肿瘤治疗领域，进展最快的微小RNA药物是miR-34a mimic的两性霉素脂质体制剂MRX34。由于miR-34a在细胞和动物水平均显示出了很好的抗肿瘤活性和安全性，美国Mirna Therapeutic公司于2013年推动了一项多中心的I期临床试验，用于治疗原发性肝癌、小细胞肺癌、淋巴瘤、黑色素瘤、多发性骨髓瘤或肾细胞癌患者，也成为第一个进入临床试验的微小RNA药物。随后，RG-101(N-乙酰-D-氨基半乳糖修饰的抗miR-122核酸片段)、RG-012(miR-21抑制剂)、RG-125/AZD4076(N-乙酰半乳糖胺修饰的miR-103/107抑制分子)、MRG-201(miR-29mimic)和MRG-106(antimiR-155锁核苷酸)等微小RNA药物相继进入临床试验，并显示出较好的疗效和安全性，已成为抗肿瘤药物研究的热点。
发明内容
有鉴于此，本发明要解决的技术问题在于提供微小RNA及其在制备抗肿瘤药物中的应用。本发明提供的微小RNA抗肿瘤效果良好。
为了实现上述发明目的，本发明提供以下技术方案：
本发明提供了RNA，包括活性链RNA和其互补链RNA；
所述活性链RNA具有如下所示的核苷酸序列中的任意一项：
I、包括22个碱基；其5’端第1位～第6位碱基序列为“UGGCAG”或第17位～第22位碱基为“UGCCCU”；或
II、具有如I所示的核苷酸序列经修饰、取代、缺失或添加一个或多个碱基获得的核苷酸序列；或
III、具有如I所示的核苷酸序列经修饰一个或多个核糖获得的核苷酸序列；或
IV、与如I所示的核苷酸序列具有至少80％同源性的序列；
所述互补为完全互补或部分互补。
在本发明的一些具体实施方案中，如I所示的核苷酸序列如SEQ ID No.1～23所示；所述其互补链RNA的序列如SEQ ID No.24～46所示。
在本发明的一些具体实施方案中，所述修饰的碱基个数为1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21或22个。
本发明还提供了转染所述RNA的宿主细胞。
在上述研究的基础上，本发明还提供了所述RNA或如权利要求4所述的宿主细胞在制备抗肿瘤药物中的应用。
在本发明的一些具体实施方案中，所述肿瘤为星形细胞瘤、间变性大细胞淋巴瘤、急性淋巴细胞白血病、急性骨髓性白血病、血管肉瘤、乳腺癌、B细胞淋巴瘤、膀胱癌、子宫颈癌、头颈癌、慢性淋巴细胞性白血病、慢性骨髓性白血病、结肠癌、直肠癌、子宫内膜癌、神经胶质瘤、胶质母细胞瘤、胃癌、胃泌素瘤、肝母细胞瘤、肝细胞癌、霍奇金淋巴瘤、卡波西肉瘤白血病、肺癌、平滑肌肉瘤、喉鳞状细胞癌、黑色素瘤、粘膜相关淋巴组织B细胞淋巴瘤、髓母细胞瘤、套细胞淋巴瘤、脑膜瘤、骨髓性白血病、多发性骨髓瘤、高危骨髓增生异常综合征、间皮瘤、神经纤维瘤、非霍奇金淋巴瘤、非小细胞肺癌、卵巢癌、食管癌、口咽癌骨肉瘤、胰腺癌、乳头状癌、前列腺癌、嗜铬细胞瘤、横纹肌肉瘤、头颈鳞状细胞癌、神经鞘瘤、小细胞肺癌、唾液腺肿瘤、散发性乳头状癌、甲状腺癌、睾丸肿瘤或尿路上皮癌。
在本发明的一些具体实施方案中，所述肿瘤的细胞包括胃癌细胞、肠癌细胞、肝癌细胞、肺癌细胞。
在本发明的一些具体实施方案中，所述肿瘤的细胞包括SGC-7901人胃癌细胞、HCT-116人肠癌细胞、HepG2人肝癌细胞、A549人肺癌细胞。
本发明还提供了一种抗肿瘤的药物，包括所述RNA或所述的宿主细胞。
在本发明的一些具体实施方案中，本发明提供的抗肿瘤的药物包括活性链和互补链；所述活性链为RNA分子或经修饰的该RNA分子，该RNA分子长度为22个碱基；其5’端第1位～第6位碱基序列为“UGGCAG”，或者第17位～第22位碱基为“UGCCCU”；所述互补链与活性链完全互补或部分互补。
本发明提供的RNA分子长度为22个碱基；其5’端第1位～第6位碱基序列为“UGGCAG”，或者第17位～第22位碱基为“UGCCCU”；采用该RNA分子能够显著抑制肿瘤细胞生长和/或增殖，从而起到抗肿瘤的作用。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍。
图1示微小RNA模拟物抑制SGC-7901人胃癌细胞生长的能力检测结果；
图2示微小RNA模拟物抑制HCT-116人肠癌细胞生长的能力检测结果；
图3示微小RNA模拟物抑制HepG2人肝癌细胞生长的能力检测结果；
图4示微小RNA模拟物抑制A549人肺癌细胞生长的能力检测结果。
具体实施方式
本发明公开了一种微小RNA及其在制备抗肿瘤药物中的应用，本领域技术人员可以借鉴本文内容，适当改进工艺参数实现。特别需要指出的是，所有类似的替换和改动对本领域技术人员来说是显而易见的，它们都被视为包括在本发明。本发明的方法及应用已经通过较佳实施例进行了描述，相关人员明显能在不脱离本发明内容、精神和范围内对本文所述的方法和应用进行改动或适当变更与组合，来实现和应用本发明技术。
本发明说明书中提及的术语定义如下：
本发明提供的RNA分子为微小RNA(microRNA，或miRNA)，其是指单链的寡核糖核酸。核糖核苷酸是由核苷酸经磷酸二酯键缩合而成长链状分子。核糖核苷酸分子由一分子碱基、一分子核糖和磷酸构成。本发明提供的miRNA的碱基有A腺嘌呤、G鸟嘌呤、C胞嘧啶、U尿嘧啶、5-甲基胞嘧啶(5-me-C)、5-羟甲基胞嘧啶、黄嘌呤、次黄嘌呤、2-氨基腺嘌呤、6-甲基和腺嘌呤和鸟嘌呤的其它烷基衍生物、腺嘌呤和鸟嘌呤的2-丙基和其它烷基衍生物、5-硫尿嘧啶和胞嘧啶、5-丙炔基尿嘧啶和嘧啶碱基的其他炔基衍生物、6-偶氮尿嘧啶、胞嘧啶和胸腺嘧啶、5-尿嘧啶(假尿嘧啶)、4-硫尿嘧啶、8-卤代、8-氨基、8-巯基、8-硫代烷基、8-羟基和其他8-取代的腺嘌呤和鸟嘌呤、5-卤素(包括5-溴、5-三氟甲基和其它5-取代的尿嘧啶和胞嘧啶)、7-甲基鸟嘌呤和7-甲基腺嘌呤、2-F-腺嘌呤、2-氨基腺嘌呤、8-氮鸟嘌呤和8-氮杂腺嘌呤、7-脱氮鸟嘌呤和7-脱氮腺嘌呤、3-脱氮鸟嘌呤和3-脱氮腺嘌呤。
所述碱基，可以是无修饰碱基，也可以是修饰碱基。
所述修饰碱基，是指碱基连接基团包括但不限于NH2、生物素、胺基、低级胺基烷基、低级烷基、NHCOCH3、乙酰基、2'-氧基-甲基(2'O-Me)、DMTO、荧光素、硫醇或吖啶。
所述修饰碱基，是指本发明提供的RNA分子链或其互补链中的1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21或22个碱基连接修饰基团。
所述修饰，是指本发明提供的RNA分子链或其互补链中的碱基连接任何一种或多种基团或其组合。
所述核糖，可以是无修饰核糖，也可以是修饰核糖。
所述修饰核糖，是指核糖连接基团包括但不限于低级烷基、烯基、炔基、烷芳基、芳烷基、O-烷芳基或O-芳烷基、SH、SCH3、Cl、Br、CN、OCN、CF3、OCF3、SOCH3、SO2CH3、ONO2、NO2、N3、NH2、杂环烷基、杂环氨基烷基、氨基烷基、聚氨基烷基、取代的甲硅烷基、RNA裂解基团、嵌入剂、用于改善微小RNA的药代动力学性质的基团或用于改善微小RNA的药效学性质的基团，以及具有相似性质的其它取代基。另外的糖取代基包括2'-O-2-甲氧基乙基(2'-O-CH2CH2OCH3)、2'-二甲基氨氧基乙氧基[2'-O-CH2-O-CH2-N(CH3)2]、烯丙基(-CH2-CH═CH2)、-O-烯丙基(-O-CH2-CH-CH2)、甲氧基(-O-CH3)、氨基丙氧基(-OCH2CH2CH2NH2)和氟(F)等。
所述修饰核糖，是指本发明提供的RNA分子链或其互补链中的任何1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21或22个核糖连接修饰基团。
所述修饰，是指本发明提供的RNA分子链或其互补链中的核糖连接任何一种或多种基团或其组合。
所述互补，又称互补配对，是指碱基通过氢键与它互补的碱基相连。互补碱基对A于U之间形成两对氢键，C与G之间形成三对氢键。
所述完全互补是指序列完全匹配且不形成粘性末端。所述部分互补是指序列完全匹配，但形成粘性末端。
本发明提供的RNA分子长度为22个碱基；其5’端第1位～第6位碱基序列为“UGGCAG”，或者第17位～第22位碱基为“UGCCCU”。
本发明还提供了与上述RNA分子序列完全互补或部分互补的RNA分子。
具体实施例中，所述部分互补形成的粘性末端为2bp。
具体实施例中，所述粘性末端位于互补RNA分子的3’端。
本发明提供的RNA分子中任意一个或多个碱基经修饰获得的RNA分子。
本发明提供的RNA分子在制备抑制肿瘤细胞生长和/或增殖的药物中的应用。
通过实验验证，本发明提供的微小RNA分子对对各种肿瘤细胞的生长均有显著的抑制作用，显示本发明提供的微小RNA分子对具有很好的广谱抗肿瘤活性。
所述肿瘤为星形细胞瘤、间变性大细胞淋巴瘤、急性淋巴细胞白血病、急性骨髓性白血病、血管肉瘤、乳腺癌、B细胞淋巴瘤、膀胱癌、子宫颈癌、头颈癌、慢性淋巴细胞性白血病、慢性骨髓性白血病、结肠癌、直肠癌、子宫内膜癌、神经胶质瘤、胶质母细胞瘤、胃癌、胃泌素瘤、肝母细胞瘤、肝细胞癌、霍奇金淋巴瘤、卡波西肉瘤白血病、肺癌、平滑肌肉瘤、喉鳞状细胞癌、黑色素瘤、粘膜相关淋巴组织B细胞淋巴瘤、髓母细胞瘤、套细胞淋巴瘤、脑膜瘤、骨髓性白血病、多发性骨髓瘤、高危骨髓增生异常综合征、间皮瘤、神经纤维瘤、非霍奇金淋巴瘤、非小细胞肺癌、卵巢癌、食管癌、口咽癌骨肉瘤、胰腺癌、乳头状癌、前列腺癌、嗜铬细胞瘤、横纹肌肉瘤、头颈鳞状细胞癌、神经鞘瘤、小细胞肺癌、唾液腺肿瘤、散发性乳头状癌、甲状腺癌、睾丸肿瘤或尿路上皮癌。
本发明进行实验的肿瘤细胞包括胃癌细胞、肠癌细胞、肝癌细胞、肺癌细胞。
具体的，进行实验的肿瘤细胞为SGC-7901人胃癌细胞、HCT-116人肠癌细胞、HepG2人肝癌细胞、A549人肺癌细胞等肿瘤细胞中的抗肿瘤活性。
本发明还提供了一种抗肿瘤的药物，包括活性链和互补链；
本发明提供的RNA分子长度为22个碱基；其5’端第1位～第6位碱基序列为“UGGCAG”，或者第17位～第22位碱基为“UGCCCU”；采用该RNA分子及其互补链能够显著抑制肿瘤细胞生长和/或增殖，从而起到抗肿瘤的作用。
本发明采用的试材皆为普通市售品，皆可于市场购得。
CCK8(东仁化学科技(上海)有限公司，日本)，微小RNA模拟物(上海吉玛基因有限公司化学合成；HPLC纯化，纯度>97％)。
电子天平(Scount SE，中国)、移液枪、高速离心机(Eppendorf，德国)、细胞培养箱、-80℃超低温冰箱(Thermo Scientific，美国)、常速离心机(Joμan，法国)、倒置显微镜(O1ympμs，日本)、电热恒温培养箱(PYX-DHS，中国)；枪尖、1.5mL EP管(Axygen，美国)、细胞培养皿及培养板、离心管(Corning，美国)。
SGC-7901人胃癌细胞、HCT-116人肠癌细胞、HepG2人肝癌细胞、A549人肺癌细胞(ATCC，美国)，细胞株经检测，无支原体污染。
本发明提供的微小RNA及其在制备抗肿瘤药物中的应用中所用原料及试剂均可由市场购得。
下面结合实施例，进一步阐述本发明：
实施例1
进行实验的RNA序列如表1：
表1微小RNA序列
本发明的实验方法包括：
1、细胞培养
所有细胞株均采用含10％胎牛血清的RPMI-1640或DMEM培养基进行培养，培养条件为37℃、5％CO2、饱和湿度。细胞生长至80％左右，用0.25％胰酶消化传代。每天观察细胞的形态及生长速度，及时更换新鲜培养基。
2、转染微小RNA
按3×103个细胞/孔的量在96孔板中接种指数生长期的细胞，培养过夜，直到细胞80％汇片。24小时后，将1、2、10、25、50、100nM微小RNA稀释至25μL不含血清和抗生素的RPMI1640培养基中，用移液枪轻轻混合均匀；将1μL脂质体2000(Invitrogen公司)悬液加入25μL不含血清和抗生素的RPMI1640培养基中，室温孵育5min；最后将两者混合在一起，充分混匀，室温静置20min，使微小RNA与脂质体充分结合。最后，将此混合物加入细胞培养孔中。
3、CCK8法检测细胞活性
向细胞培养孔中加入CCK-8溶液，在培养箱中继续培养0.5小时后，酶标仪检测450nm处吸光值(A)，计算抑制率＝(样品A值-空白A值)/(对照A值-空白A值)×100％。
本发明的实验结果为：
测定如SEQ ID NO.1～23所示的RNA活性链和如SEQ ID NO.24～46所示的RNA互补链抑制SGC-7901人胃癌细胞、HCT-116人肠癌细胞、HepG2人肝癌细胞、A549人肺癌细胞生长的活性。结果如图1-4和表2所示，各种肿瘤细胞的生长均被显著抑制，显示这些微小RNA具有很好的广谱抗肿瘤活性。
表2微小RNA抑制肿瘤细胞生长的IC50值(nM)
注：“无”表示细胞不敏感。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
序列表
<110> 苏州大学
<120> 微小RNA及其在制备抗肿瘤药物中的应用
<130> MP1905264
<160> 46
<170> SIPOSequenceListing 1.0
<210> 1
<211> 14
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 1
ggcagggcgg acgc 14
<210> 2
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 2
uggcaggugg cgucguugua ac 22
<210> 4
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 4
uggcagaaau ggacaccacg ac 22
<210> 5
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 5
uggcagggga guugaacuug uc 22
<210> 6
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 6
uggcagcuca ucggauuucu uu 22
<210> 7
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 7
uggcagauaa aagagauauu cu 22
<210> 8
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 8
uggcagcgcu ccucccucuu ua 22
<210> 9
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 9
uggcaguacg guccauccaa ug 22
<210> 10
<211> 17
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 10
ggcaggccac acaggcc 17
<210> 11
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 11
uggcagcccu aaacuugaau ag 22
<210> 12
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 12
uggcagaaua gaauugcugc ua 22
<210> 13
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 13
uggcagagcu uuaccgugcg uu 22
<210> 14
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 14
cggccucaug ucggauugcc cu 22
<210> 15
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 15
ggugaguaga agaaugugcc cu 22
<210> 16
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 16
ccagccccau guccacugcc cu 22
<210> 17
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 17
uucaagauga aauuugugcc cu 22
<210> 18
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 18
aaucuuuuau uuugagugcc cu 22
<210> 19
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 19
aacucuucca aauccaugcc cu 22
<210> 20
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 20
ggacuuguua gaauguugcc cu 22
<210> 21
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 21
uauuguaaug acauguugcc cu 22
<210> 22
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 22
ugcauagcau cuagguugcc cu 22
<210> 23
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 23
agccacaaga auucugugcc cu 22
<210> 24
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 24
aagggggcuc ccccucugcc cu 22
<210> 25
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 25
gcaagucacg ccaacugcca uu 22
<210> 26
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 26
uacaacgacg ccaccugcca uu 22
<210> 27
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 27
cguggugucc auuucugcca uu 22
<210> 28
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 28
caaguucaac uccccugcca uu 22
<210> 29
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 29
agaaauccga ugagcugcca uu 22
<210> 30
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 30
aauaucucuu uuaucugcca uu 22
<210> 31
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 31
aagagggagg agcgcugcca uu 22
<210> 32
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 32
uuggauggac cguacugcca uu 22
<210> 33
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 33
ccaugaaaug uggccugcca uu 22
<210> 34
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 34
auucaaguuu agggcugcca uu 22
<210> 35
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 35
gcagcaauuc uauucugcca uu 22
<210> 36
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 36
cgcacgguaa agcucugcca uu 22
<210> 37
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 37
ggcaauccga caugaggccg uu 22
<210> 38
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 38
ggcacauucu ucuacucacc uu 22
<210> 39
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 39
ggcaguggac auggggcugg uu 22
<210> 40
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 40
ggcacaaauu ucaucuugaa uu 22
<210> 41
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 41
ggcacucaaa auaaaagauu uu 22
<210> 42
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 42
ggcauggauu uggaagaguu uu 22
<210> 43
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 43
ggcaacauuc uaacaagucc uu 22
<210> 44
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 44
ggcaacaugu cauuacaaua uu 22
<210> 44
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 44
ggcaaccuag augcuaugca uu 22
<210> 45
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 45
ggcacagaau ucuuguggcu uu 22
<210> 46
<211> 22
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 46
ggcagagggg gagcccccuu uu 22
Claims (3)
1.RNA，其特征在于，包括活性链RNA和其互补链RNA；所述活性链RNA的核苷酸序列如SEQ ID No.18所示，其互补链RNA的核苷酸序列如SEQ ID No.41所示。
2.如权利要求1所述RNA在制备抗肿瘤药物中的应用，所述肿瘤为胃癌。
3.一种抗肿瘤的药物，其特征在于，包括如权利要求1所述RNA；
所述肿瘤为胃癌。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911293658.5A CN110904108B (zh) | 2019-12-16 | 2019-12-16 | 微小rna及其在制备抗肿瘤药物中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911293658.5A CN110904108B (zh) | 2019-12-16 | 2019-12-16 | 微小rna及其在制备抗肿瘤药物中的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110904108A CN110904108A (zh) | 2020-03-24 |
| CN110904108B true CN110904108B (zh) | 2024-01-30 |
Family
ID=69825930
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911293658.5A Active CN110904108B (zh) | 2019-12-16 | 2019-12-16 | 微小rna及其在制备抗肿瘤药物中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110904108B (zh) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2007299748A1 (en) * | 2006-09-19 | 2008-03-27 | Asuragen, Inc. | miR-15, miR-26, miR -31,miR -145, miR-147, miR-188, miR-215, miR-216 miR-331, mmu-miR-292-3p regulated genes and pathways as targets for therapeutic intervention |
| CN107699565A (zh) * | 2017-11-24 | 2018-02-16 | 苏州大学 | 微小rna及其在制备抗肿瘤药物中的应用 |
-
2019
- 2019-12-16 CN CN201911293658.5A patent/CN110904108B/zh active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2007299748A1 (en) * | 2006-09-19 | 2008-03-27 | Asuragen, Inc. | miR-15, miR-26, miR -31,miR -145, miR-147, miR-188, miR-215, miR-216 miR-331, mmu-miR-292-3p regulated genes and pathways as targets for therapeutic intervention |
| CN107699565A (zh) * | 2017-11-24 | 2018-02-16 | 苏州大学 | 微小rna及其在制备抗肿瘤药物中的应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110904108A (zh) | 2020-03-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP2261333B1 (en) | Pharmaceutical composition comprising anti-miRNA antisense oligonucleotides | |
| JP6010458B2 (ja) | 非対称二本鎖ｒｎａによる特異的阻害のための、方法および組成物 | |
| EP2203559B1 (en) | Combination treatment for the treatment of hepatitis c virus infection | |
| JP5198430B2 (ja) | ａｎｔｉｍｉＲＮＡアンチセンスオリゴヌクレオチドを含む医薬組成物 | |
| JP5976643B2 (ja) | 二本鎖ｒｎａによるベータ−カテニンの特異的阻害に対する方法と組成物 | |
| JP2018507866A (ja) | マイクロｒｎａを有効成分として含む癌治療用医薬組成物 | |
| US20230348911A1 (en) | Antisense oligonucleotides for modulation of long noncoding rnas | |
| JP2018500027A (ja) | リガンド修飾二本鎖核酸 | |
| KR20080068019A (ko) | 아포지단백질-ｂ100 발현 억제용 ｒｎａ 길항제 화합물 | |
| US20100047909A1 (en) | Nucleic acid compounds for inhibiting vegf family gene expression and uses thereof | |
| Detich et al. | A conserved 3′-untranslated element mediates growth regulation of DNA methyltransferase 1 and inhibits its transforming activity | |
| Liu et al. | MiR-542-3p exerts tumor suppressive functions in non-small cell lung cancer cells by upregulating FTSJ2 | |
| US20100055782A1 (en) | Nucleic acid compounds for inhibiting myc gene expression and uses thereof | |
| CN101437942A (zh) | 包含抗微小rna反义寡核苷酸的药物组合物 | |
| CN107699565B (zh) | 微小rna及其在制备抗肿瘤药物中的应用 | |
| CN110904108B (zh) | 微小rna及其在制备抗肿瘤药物中的应用 | |
| CN107709561B (zh) | 修饰的siRNA及含有该修饰的siRNA的药物组合物 | |
| EP2716758A1 (en) | OLIGONUCLEOTIDE DERIVATIVE, OLIGONUCLEOTIDE DERIVATIVE-CONTAINING PHARMACEUTICAL COMPOSITION FOR TREATMENT AND PHARMACEUTICAL COMPOSITION FOR DIAGNOSIS, AND OLIGONUCLEOTIDE DERIVATIVE FOR REGULATION OF miRNA FUNCTION | |
| CN106581676B (zh) | 癌症标志物、治疗癌症的药物组分及用途 | |
| JP2022120380A (ja) | ｍｉＲＮＡ１３３－ｂ誘導体及びその利用 | |
| CN108192895B (zh) | 靶向NOB1基因的siRNA分子及其应用 | |
| CN114591960A (zh) | 长反义rna诱导细胞干扰素生成、抑制细胞生长的方法 | |
| AU2012216487B2 (en) | Pharmaceutical composition comprising anti-miRNA antisense oligonucleotides | |
| KR20120116766A (ko) | Ａｄｅｎｉｎｅ ｎｕｃｌｅｏｔｉｄｅ ｔｒａｎｓｌｏｃａｔｏｒ ２ 발현 또는 활성 억제제를 이용한 간암 치료 방법 | |
| Thomas | Syntheses and applications of small molecule inhibitors of miRNAs miR-21 and miR-122 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |