CN111812309A - 肿瘤骨转移的尿液蛋白标记物及其用途 - Google Patents
肿瘤骨转移的尿液蛋白标记物及其用途 Download PDFInfo
- Publication number
- CN111812309A CN111812309A CN201910289068.9A CN201910289068A CN111812309A CN 111812309 A CN111812309 A CN 111812309A CN 201910289068 A CN201910289068 A CN 201910289068A CN 111812309 A CN111812309 A CN 111812309A
- Authority
- CN
- China
- Prior art keywords
- protein
- alpha
- chain
- bone metastasis
- adhesion molecule
- 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.)
- Pending
Links
- 210000000988 bone and bone Anatomy 0.000 title claims abstract description 45
- 210000002700 urine Anatomy 0.000 title claims abstract description 41
- 206010027476 Metastases Diseases 0.000 title claims abstract description 39
- 230000009401 metastasis Effects 0.000 title claims abstract description 39
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 34
- 239000012474 protein marker Substances 0.000 title abstract description 7
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 78
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 78
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 21
- 108060008226 thioredoxin Proteins 0.000 claims abstract description 15
- 229940094937 thioredoxin Drugs 0.000 claims abstract description 15
- XJOTXKZIRSHZQV-RXHOOSIZSA-N (3S)-3-amino-4-[[(2S,3R)-1-[[(2S)-1-[[(2S)-1-[(2S)-2-[[(2S,3S)-1-[[(1R,6R,12R,17R,20S,23S,26R,31R,34R,39R,42S,45S,48S,51S,59S)-51-(4-aminobutyl)-31-[[(2S)-6-amino-1-[[(1S,2R)-1-carboxy-2-hydroxypropyl]amino]-1-oxohexan-2-yl]carbamoyl]-20-benzyl-23-[(2S)-butan-2-yl]-45-(3-carbamimidamidopropyl)-48-(hydroxymethyl)-42-(1H-imidazol-4-ylmethyl)-59-(2-methylsulfanylethyl)-7,10,19,22,25,33,40,43,46,49,52,54,57,60,63,64-hexadecaoxo-3,4,14,15,28,29,36,37-octathia-8,11,18,21,24,32,41,44,47,50,53,55,58,61,62,65-hexadecazatetracyclo[32.19.8.26,17.212,39]pentahexacontan-26-yl]amino]-3-methyl-1-oxopentan-2-yl]carbamoyl]pyrrolidin-1-yl]-1-oxo-3-phenylpropan-2-yl]amino]-3-(1H-imidazol-4-yl)-1-oxopropan-2-yl]amino]-3-hydroxy-1-oxobutan-2-yl]amino]-4-oxobutanoic acid Chemical compound CC[C@H](C)[C@H](NC(=O)[C@@H]1CCCN1C(=O)[C@H](Cc1ccccc1)NC(=O)[C@H](Cc1cnc[nH]1)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(O)=O)[C@@H](C)O)C(=O)N[C@H]1CSSC[C@H](NC(=O)[C@@H]2CSSC[C@@H]3NC(=O)[C@@H]4CSSC[C@H](NC(=O)[C@H](Cc5ccccc5)NC(=O)[C@@H](NC1=O)[C@@H](C)CC)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](Cc1cnc[nH]1)NC3=O)C(=O)NCC(=O)N[C@@H](CCSC)C(=O)N2)C(=O)NCC(=O)N4)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XJOTXKZIRSHZQV-RXHOOSIZSA-N 0.000 claims abstract description 8
- 101710102159 Atrial natriuretic peptide receptor 2 Proteins 0.000 claims abstract description 8
- 102100039341 Atrial natriuretic peptide receptor 2 Human genes 0.000 claims abstract description 8
- 102100040510 Galectin-3-binding protein Human genes 0.000 claims abstract description 8
- 101710197901 Galectin-3-binding protein Proteins 0.000 claims abstract description 8
- 108010041834 Growth Differentiation Factor 15 Proteins 0.000 claims abstract description 8
- 102000018697 Membrane Proteins Human genes 0.000 claims abstract description 8
- 108010052285 Membrane Proteins Proteins 0.000 claims abstract description 8
- 108010061952 Orosomucoid Proteins 0.000 claims abstract description 8
- 102000012404 Orosomucoid Human genes 0.000 claims abstract description 8
- 102000004264 Osteopontin Human genes 0.000 claims abstract description 8
- 108010081689 Osteopontin Proteins 0.000 claims abstract description 8
- 108010027007 Uromodulin Proteins 0.000 claims abstract description 8
- 108010081355 beta 2-Microglobulin Proteins 0.000 claims abstract description 8
- 102000015736 beta 2-Microglobulin Human genes 0.000 claims abstract description 8
- 102000018511 hepcidin Human genes 0.000 claims abstract description 8
- 108060003558 hepcidin Proteins 0.000 claims abstract description 8
- 229940066919 hepcidin Drugs 0.000 claims abstract description 8
- 230000001105 regulatory effect Effects 0.000 claims description 16
- 102000002933 Thioredoxin Human genes 0.000 claims description 14
- 102100033448 Lysosomal alpha-glucosidase Human genes 0.000 claims description 12
- 108010028144 alpha-Glucosidases Proteins 0.000 claims description 12
- 238000011002 quantification Methods 0.000 claims description 11
- 102100038838 2-Hydroxyacid oxidase 2 Human genes 0.000 claims description 7
- 101710160335 2-Hydroxyacid oxidase 2 Proteins 0.000 claims description 7
- 102000001762 6-phosphogluconolactonase Human genes 0.000 claims description 7
- 108010029731 6-phosphogluconolactonase Proteins 0.000 claims description 7
- 101710182094 8-oxo-dGTP diphosphatase Proteins 0.000 claims description 7
- 102100022252 A-kinase anchor protein SPHKAP Human genes 0.000 claims description 7
- 108010022579 ATP dependent 26S protease Proteins 0.000 claims description 7
- 102100023619 ATP synthase F(0) complex subunit B1, mitochondrial Human genes 0.000 claims description 7
- 101710201407 ATP synthase F(0) complex subunit B1, mitochondrial Proteins 0.000 claims description 7
- 102000005606 Activins Human genes 0.000 claims description 7
- 108010059616 Activins Proteins 0.000 claims description 7
- 108020004306 Alpha-ketoglutarate dehydrogenase Proteins 0.000 claims description 7
- 102000006589 Alpha-ketoglutarate dehydrogenase Human genes 0.000 claims description 7
- 102000008102 Ankyrins Human genes 0.000 claims description 7
- 108010049777 Ankyrins Proteins 0.000 claims description 7
- 102100037320 Apolipoprotein A-IV Human genes 0.000 claims description 7
- 108010076807 Apolipoprotein C-I Proteins 0.000 claims description 7
- 102000011772 Apolipoprotein C-I Human genes 0.000 claims description 7
- 108010074051 C-Reactive Protein Proteins 0.000 claims description 7
- 102100032752 C-reactive protein Human genes 0.000 claims description 7
- 102100029390 CMRF35-like molecule 1 Human genes 0.000 claims description 7
- 101710157060 CMRF35-like molecule 1 Proteins 0.000 claims description 7
- 102100027473 Cartilage oligomeric matrix protein Human genes 0.000 claims description 7
- 101710176668 Cartilage oligomeric matrix protein Proteins 0.000 claims description 7
- 108090000613 Cathepsin S Proteins 0.000 claims description 7
- 102100035654 Cathepsin S Human genes 0.000 claims description 7
- 108010072135 Cell Adhesion Molecule-1 Proteins 0.000 claims description 7
- 102100024649 Cell adhesion molecule 1 Human genes 0.000 claims description 7
- 101710144787 Cell surface glycoprotein MUC18 Proteins 0.000 claims description 7
- 102100023126 Cell surface glycoprotein MUC18 Human genes 0.000 claims description 7
- 108010028778 Complement C4 Proteins 0.000 claims description 7
- 102100024339 Complement component C6 Human genes 0.000 claims description 7
- 101710203188 Complement component C6 Proteins 0.000 claims description 7
- 102100030012 Deoxyribonuclease-1 Human genes 0.000 claims description 7
- 101710206036 Deoxyribonuclease-1 Proteins 0.000 claims description 7
- 108010028196 Dihydropteridine Reductase Proteins 0.000 claims description 7
- 102100022317 Dihydropteridine reductase Human genes 0.000 claims description 7
- 102000016680 Dioxygenases Human genes 0.000 claims description 7
- 108010028143 Dioxygenases Proteins 0.000 claims description 7
- 102100020751 Dipeptidyl peptidase 2 Human genes 0.000 claims description 7
- 101710087084 Dipeptidyl peptidase 2 Proteins 0.000 claims description 7
- 108010080865 Factor XII Proteins 0.000 claims description 7
- 102000000429 Factor XII Human genes 0.000 claims description 7
- 102000030914 Fatty Acid-Binding Human genes 0.000 claims description 7
- 101710137044 Fibrinogen alpha chain Proteins 0.000 claims description 7
- 102000016359 Fibronectins Human genes 0.000 claims description 7
- 108010067306 Fibronectins Proteins 0.000 claims description 7
- 102000016970 Follistatin Human genes 0.000 claims description 7
- 108010014612 Follistatin Proteins 0.000 claims description 7
- 102100021023 Gamma-glutamyl hydrolase Human genes 0.000 claims description 7
- 102000004878 Gelsolin Human genes 0.000 claims description 7
- 108090001064 Gelsolin Proteins 0.000 claims description 7
- 102000006587 Glutathione peroxidase Human genes 0.000 claims description 7
- 108700016172 Glutathione peroxidases Proteins 0.000 claims description 7
- 102100040896 Growth/differentiation factor 15 Human genes 0.000 claims description 7
- 101710154606 Hemagglutinin Proteins 0.000 claims description 7
- 102000008949 Histocompatibility Antigens Class I Human genes 0.000 claims description 7
- 108010088652 Histocompatibility Antigens Class I Proteins 0.000 claims description 7
- 101000825204 Homo sapiens A-kinase anchor protein SPHKAP Proteins 0.000 claims description 7
- 102100039283 Hyaluronidase-1 Human genes 0.000 claims description 7
- 101710199679 Hyaluronidase-1 Proteins 0.000 claims description 7
- 101710187342 Ig lambda-2 chain C region Proteins 0.000 claims description 7
- 102100029620 Immunoglobulin lambda constant 2 Human genes 0.000 claims description 7
- 102100023350 Integral membrane protein 2B Human genes 0.000 claims description 7
- 101710180845 Integral membrane protein 2B Proteins 0.000 claims description 7
- 102000015271 Intercellular Adhesion Molecule-1 Human genes 0.000 claims description 7
- 108010064593 Intercellular Adhesion Molecule-1 Proteins 0.000 claims description 7
- 108010015372 Low Density Lipoprotein Receptor-Related Protein-2 Proteins 0.000 claims description 7
- 102100021922 Low-density lipoprotein receptor-related protein 2 Human genes 0.000 claims description 7
- 101710094503 Metallothionein-1 Proteins 0.000 claims description 7
- 101710093908 Outer capsid protein VP4 Proteins 0.000 claims description 7
- 101710135467 Outer capsid protein sigma-1 Proteins 0.000 claims description 7
- 102100039792 Oxidized purine nucleoside triphosphate hydrolase Human genes 0.000 claims description 7
- 101710169326 Oxidized purine nucleoside triphosphate hydrolase Proteins 0.000 claims description 7
- 102000005327 Palmitoyl protein thioesterase Human genes 0.000 claims description 7
- 108020002591 Palmitoyl protein thioesterase Proteins 0.000 claims description 7
- 108010050808 Procollagen Proteins 0.000 claims description 7
- 102000058035 Prostaglandin reductase 2 Human genes 0.000 claims description 7
- 108700001195 Prostaglandin reductase 2 Proteins 0.000 claims description 7
- 101710176177 Protein A56 Proteins 0.000 claims description 7
- 102100027378 Prothrombin Human genes 0.000 claims description 7
- 108010094028 Prothrombin Proteins 0.000 claims description 7
- 102100028688 Putative glycosylation-dependent cell adhesion molecule 1 Human genes 0.000 claims description 7
- 102100036164 Putative phospholipase B-like 2 Human genes 0.000 claims description 7
- 101710200837 Putative phospholipase B-like 2 Proteins 0.000 claims description 7
- 101710141795 Ribonuclease inhibitor Proteins 0.000 claims description 7
- 102100037968 Ribonuclease inhibitor Human genes 0.000 claims description 7
- 229940122208 Ribonuclease inhibitor Drugs 0.000 claims description 7
- 101710103121 Sodium/glucose cotransporter 1 Proteins 0.000 claims description 7
- 102100020885 Sodium/glucose cotransporter 1 Human genes 0.000 claims description 7
- 102100038836 Superoxide dismutase [Cu-Zn] Human genes 0.000 claims description 7
- 101710139715 Superoxide dismutase [Cu-Zn] Proteins 0.000 claims description 7
- 101710187830 Tumor necrosis factor receptor superfamily member 1B Proteins 0.000 claims description 7
- 102100033733 Tumor necrosis factor receptor superfamily member 1B Human genes 0.000 claims description 7
- 102100040613 Uromodulin Human genes 0.000 claims description 7
- 102000050760 Vitamin D-binding protein Human genes 0.000 claims description 7
- 101710179590 Vitamin D-binding protein Proteins 0.000 claims description 7
- 108050001627 Voltage-dependent anion-selective channel protein 1 Proteins 0.000 claims description 7
- 102000011404 Voltage-dependent anion-selective channel protein 1 Human genes 0.000 claims description 7
- 239000000488 activin Substances 0.000 claims description 7
- 108010073614 apolipoprotein A-IV Proteins 0.000 claims description 7
- 210000004556 brain Anatomy 0.000 claims description 7
- 238000003745 diagnosis Methods 0.000 claims description 7
- 108091022862 fatty acid binding Proteins 0.000 claims description 7
- 108010062699 gamma-Glutamyl Hydrolase Proteins 0.000 claims description 7
- 239000000185 hemagglutinin Substances 0.000 claims description 7
- 108010037896 heparin-binding hemagglutinin Proteins 0.000 claims description 7
- 229940039716 prothrombin Drugs 0.000 claims description 7
- 239000003161 ribonuclease inhibitor Substances 0.000 claims description 7
- 108010012704 sulfated glycoprotein p50 Proteins 0.000 claims description 7
- 102100039087 Peptidyl-alpha-hydroxyglycine alpha-amidating lyase Human genes 0.000 claims description 6
- 108091000080 Phosphotransferase Proteins 0.000 claims description 6
- 238000013399 early diagnosis Methods 0.000 claims description 6
- 108010007262 peptidylglycine monooxygenase Proteins 0.000 claims description 6
- 102000020233 phosphotransferase Human genes 0.000 claims description 6
- 102100029767 Copper transport protein ATOX1 Human genes 0.000 claims description 5
- 101710120376 Copper transport protein ATOX1 Proteins 0.000 claims description 5
- 102100030497 Cytochrome c Human genes 0.000 claims description 5
- 108010075031 Cytochromes c Proteins 0.000 claims description 5
- 102000004357 Transferases Human genes 0.000 claims description 5
- 108090000992 Transferases Proteins 0.000 claims description 5
- 239000008187 granular material Substances 0.000 claims description 5
- 102000004157 Hydrolases Human genes 0.000 claims description 4
- 108090000604 Hydrolases Proteins 0.000 claims description 4
- 102100031357 L-lactate dehydrogenase C chain Human genes 0.000 claims description 4
- 101710111525 L-lactate dehydrogenase C chain Proteins 0.000 claims description 4
- 230000002438 mitochondrial effect Effects 0.000 claims description 4
- 102000003855 L-lactate dehydrogenase Human genes 0.000 claims description 3
- 108700023483 L-lactate dehydrogenases Proteins 0.000 claims description 3
- 239000000427 antigen Substances 0.000 claims description 3
- 108091007433 antigens Proteins 0.000 claims description 3
- 102000036639 antigens Human genes 0.000 claims description 3
- 230000003247 decreasing effect Effects 0.000 claims description 3
- 239000012634 fragment Substances 0.000 claims description 3
- KPGXRSRHYNQIFN-UHFFFAOYSA-N 2-oxoglutaric acid Chemical compound OC(=O)CCC(=O)C(O)=O KPGXRSRHYNQIFN-UHFFFAOYSA-N 0.000 claims description 2
- 108010052832 Cytochromes Proteins 0.000 claims description 2
- 102000018832 Cytochromes Human genes 0.000 claims description 2
- 102100031920 Dihydrolipoyllysine-residue succinyltransferase component of 2-oxoglutarate dehydrogenase complex, mitochondrial Human genes 0.000 claims description 2
- 108030000716 Dihydrolipoyllysine-residue succinyltransferases Proteins 0.000 claims description 2
- 108060003393 Granulin Proteins 0.000 claims description 2
- 239000004472 Lysine Substances 0.000 claims description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 2
- 102000017941 granulin Human genes 0.000 claims description 2
- 230000002132 lysosomal effect Effects 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- YIEDSISPYKQADU-UHFFFAOYSA-N n-acetyl-n-[2-methyl-4-[(2-methylphenyl)diazenyl]phenyl]acetamide Chemical compound C1=C(C)C(N(C(C)=O)C(=O)C)=CC=C1N=NC1=CC=CC=C1C YIEDSISPYKQADU-UHFFFAOYSA-N 0.000 claims description 2
- 239000008194 pharmaceutical composition Substances 0.000 claims description 2
- 230000003248 secreting effect Effects 0.000 claims description 2
- 230000000392 somatic effect Effects 0.000 claims description 2
- 238000003860 storage Methods 0.000 claims description 2
- 102100031752 Fibrinogen alpha chain Human genes 0.000 claims 3
- 102000008109 Mixed Function Oxygenases Human genes 0.000 claims 1
- 108010074633 Mixed Function Oxygenases Proteins 0.000 claims 1
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 229920001192 peptidylglycine Polymers 0.000 claims 1
- 238000012216 screening Methods 0.000 abstract description 5
- 238000010171 animal model Methods 0.000 abstract description 3
- 108010001387 Fatty Acid-Binding Protein 7 Proteins 0.000 abstract 1
- 102000002140 Fatty Acid-Binding Protein 7 Human genes 0.000 abstract 1
- 102000000597 Growth Differentiation Factor 15 Human genes 0.000 abstract 1
- 102000004316 Oxidoreductases Human genes 0.000 abstract 1
- 108090000854 Oxidoreductases Proteins 0.000 abstract 1
- 102100036407 Thioredoxin Human genes 0.000 abstract 1
- 102000018614 Uromodulin Human genes 0.000 abstract 1
- 230000000295 complement effect Effects 0.000 abstract 1
- 238000001514 detection method Methods 0.000 abstract 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 abstract 1
- 239000003550 marker Substances 0.000 abstract 1
- -1 mitochondria Proteins 0.000 abstract 1
- 210000003470 mitochondria Anatomy 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 description 50
- 241000700159 Rattus Species 0.000 description 24
- 210000004027 cell Anatomy 0.000 description 11
- 238000000034 method Methods 0.000 description 11
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- 108010019160 Pancreatin Proteins 0.000 description 6
- 201000010099 disease Diseases 0.000 description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 6
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 6
- 102000007079 Peptide Fragments Human genes 0.000 description 5
- 108010033276 Peptide Fragments Proteins 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 150000002500 ions Chemical class 0.000 description 5
- 229940055695 pancreatin Drugs 0.000 description 5
- 238000001228 spectrum Methods 0.000 description 5
- 210000002303 tibia Anatomy 0.000 description 5
- 102400000524 Fibrinogen alpha chain Human genes 0.000 description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- 239000000090 biomarker Substances 0.000 description 4
- 201000011510 cancer Diseases 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000003384 imaging method Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 108090000765 processed proteins & peptides Proteins 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 102100024647 Protein ABHD14B Human genes 0.000 description 3
- 101710173275 Protein ABHD14B Proteins 0.000 description 3
- 108010029485 Protein Isoforms Proteins 0.000 description 3
- 102000001708 Protein Isoforms Human genes 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 235000019253 formic acid Nutrition 0.000 description 3
- 230000004927 fusion Effects 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 208000031648 Body Weight Changes Diseases 0.000 description 2
- 101000727865 Homo sapiens Copper transport protein ATOX1 Proteins 0.000 description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 2
- 208000002193 Pain Diseases 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 230000004579 body weight change Effects 0.000 description 2
- 230000037182 bone density Effects 0.000 description 2
- 210000001185 bone marrow Anatomy 0.000 description 2
- 230000004097 bone metabolism Effects 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000012869 ethanol precipitation Methods 0.000 description 2
- 238000013467 fragmentation Methods 0.000 description 2
- 238000006062 fragmentation reaction Methods 0.000 description 2
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 2
- 201000005202 lung cancer Diseases 0.000 description 2
- 208000020816 lung neoplasm Diseases 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 238000000465 moulding Methods 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 238000004445 quantitative analysis Methods 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 230000002485 urinary effect Effects 0.000 description 2
- ATRRKUHOCOJYRX-UHFFFAOYSA-N Ammonium bicarbonate Chemical compound [NH4+].OC([O-])=O ATRRKUHOCOJYRX-UHFFFAOYSA-N 0.000 description 1
- 229910000013 Ammonium bicarbonate Inorganic materials 0.000 description 1
- 208000006386 Bone Resorption Diseases 0.000 description 1
- 206010005949 Bone cancer Diseases 0.000 description 1
- 208000018084 Bone neoplasm Diseases 0.000 description 1
- 238000009010 Bradford assay Methods 0.000 description 1
- 208000014644 Brain disease Diseases 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 206010006895 Cachexia Diseases 0.000 description 1
- 206010058019 Cancer Pain Diseases 0.000 description 1
- 201000000274 Carcinosarcoma Diseases 0.000 description 1
- 206010017076 Fracture Diseases 0.000 description 1
- 208000037147 Hypercalcaemia Diseases 0.000 description 1
- 206010027452 Metastases to bone Diseases 0.000 description 1
- 206010033892 Paraplegia Diseases 0.000 description 1
- 108090000244 Rat Proteins Proteins 0.000 description 1
- 241000700157 Rattus norvegicus Species 0.000 description 1
- RPDSFWLVWFJAHW-UHFFFAOYSA-N acetamide;hydroiodide Chemical compound I.CC(N)=O RPDSFWLVWFJAHW-UHFFFAOYSA-N 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 230000002152 alkylating effect Effects 0.000 description 1
- 235000012538 ammonium bicarbonate Nutrition 0.000 description 1
- 239000001099 ammonium carbonate Substances 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 238000010256 biochemical assay Methods 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 230000024279 bone resorption Effects 0.000 description 1
- 238000007623 carbamidomethylation reaction Methods 0.000 description 1
- 238000013375 chromatographic separation Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000007071 enzymatic hydrolysis Effects 0.000 description 1
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 230000009097 homeostatic mechanism Effects 0.000 description 1
- 230000000148 hypercalcaemia Effects 0.000 description 1
- 208000030915 hypercalcemia disease Diseases 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 210000002414 leg Anatomy 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 210000005075 mammary gland Anatomy 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000001394 metastastic effect Effects 0.000 description 1
- 206010061289 metastatic neoplasm Diseases 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004660 morphological change Effects 0.000 description 1
- 238000013188 needle biopsy Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 229960001412 pentobarbital Drugs 0.000 description 1
- WEXRUCMBJFQVBZ-UHFFFAOYSA-N pentobarbital Chemical compound CCCC(C)C1(CC)C(=O)NC(=O)NC1=O WEXRUCMBJFQVBZ-UHFFFAOYSA-N 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 125000004805 propylene group Chemical group [H]C([H])([H])C([H])([*:1])C([H])([H])[*:2] 0.000 description 1
- 238000011552 rat model Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 238000002603 single-photon emission computed tomography Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000013222 sprague-dawley male rat Methods 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000004885 tandem mass spectrometry Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 230000008646 thermal stress Effects 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 239000000165 urinary protein biomarker Substances 0.000 description 1
- 208000014001 urinary system disease Diseases 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5091—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing the pathological state of an organism
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
- G01N30/7233—Mass spectrometers interfaced to liquid or supercritical fluid chromatograph
- G01N30/724—Nebulising, aerosol formation or ionisation
- G01N30/7266—Nebulising, aerosol formation or ionisation by electric field, e.g. electrospray
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6848—Methods of protein analysis involving mass spectrometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/60—Complex ways of combining multiple protein biomarkers for diagnosis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/70—Mechanisms involved in disease identification
- G01N2800/7023—(Hyper)proliferation
- G01N2800/7028—Cancer
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Bioinformatics & Computational Biology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physiology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Dispersion Chemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
本申请涉及肿瘤骨转移的尿液蛋白标记物及其用途。本申请涉及选自以下的尿液蛋白标记物在预测肿瘤骨转移风险和/或用于诊断肿瘤骨转移中的用途：心房利钠肽受体2、半乳糖凝集素‑3结合蛋白、铁调素、骨桥蛋白、生长分化因子15、羟基酸氧化酶2、NKG2‑DII型整合膜蛋白、脂肪酸结合蛋白，脑、补体成分C6、α1酸性糖蛋白、硫氧还蛋白，线粒体、β‑2‑微球蛋白、尿调节素等。本申请建立了筛选与肿瘤骨转移相关的尿液蛋白标记物的动物模型，使用上述建立的动物模型获得与肿瘤骨转移相关的尿液蛋白标记物，以及标记物的检测试剂在制备诊断受试者肿瘤骨转移的试剂盒中的用途。
Description
技术领域
本发明涉及临床医学领域；具体而言，本发明涉及肿瘤骨转移相关的尿液蛋白标记物。具体而言，本发明涉及利用肿瘤骨转移的大鼠模型和质谱分析尿液蛋白组学技术获得与人肿瘤骨转移相关的尿液蛋白标记物，以及所述尿液蛋白标记物的用途。
背景技术
骨骼是恶性肿瘤常见的转移部位，仅此于肺、肝。数据表明有75％至90％的晚期肿瘤患者发生骨转移且经历显著的癌症疼痛，参见Montiel-Ruiz RM等人Bone cancer pain:from preclinical pharmacology to clinical trials.Gac Med Mex,2013,149(2)：204-211。
肿瘤骨转移不仅影响患者的生活质量，同时也导致患者遭受贫血、骨折、截瘫、高血钙、疼痛和恶病质等痛苦，增加死亡率，参见Coleman,R.E.Skeletal complications ofmalignancy.Cancer,80,1588–1594。因此，肿瘤骨转移的早期诊治对于提高患者的生活质量、延长生存率，具有重大意义。
穿刺活检是诊断骨转移的金标准，但由于其为有创检查，临床上依从性较低。影像学检查在骨转移的筛查和早期诊断中发挥着不可替代的作用，诊断骨转移的影像学检查方法主要有普通X射线平片、CT、MRI、全身骨显像(WBS)、SPECT、PE/CT等。但存在缺陷，其早期诊断并不理想，发现时多已发病，参见孙燕等，肺癌骨转移诊疗专家共识2014版.中国肺癌杂志,2014,95(2):57-72；和Clamp,A等人2004.Assessment of therapeutic response inpatients with metastatic bone disease.Lancet Oncology,5(10),607–616。骨转移发生时，骨代谢生化指标的变化明显早于影像学所发现的形态学改变，研究者们开始关注骨代谢指标，寻找疾病标志物，参见Stella D'Oronzo等人The value of biomarkers inbone metastasis[J].European Journal of Cancer Care,2017,26(6)。所以亟需找到一种无创、简单、高敏感性、高特异性的生物学标记物。
尿液是机体代谢所产生的排泄废物，不受内环境稳态机制的调控，因此尿液可以更加灵敏、早期地反应机体的生理病理变化，即尿液是比血液更好的寻找疾病标志物的重要来源，参见Gao Y.Urine-an untapped goldmine for biomarker discovery.Sci ChinaLife Sci,2013,56:1145-1146；和Wu J,Gao Y.Physiological conditions can bereflected in human urine proteome and metabolome.Expert review of proteomics,2015,12:623-636；和Li M,Zhao M,Gao Y.Changes of proteins induced byanticoagulants can be more sensitively detected in urine than in plasma.SciChina Life Sci,2014,57:649-654。
此外，尿液还具有可大量、连续性、无创性收集的优点。据Urinary ProteinBiomarker Database(6/29/2017更新)，有924个尿液候选蛋白标记物被报道与159种疾病相关，参见Shao C等人A tool for biomarker discovery in the urinary proteome:Amanually curated human and animal urine protein biomarker database.Mol CellProteomics,2011,10。除泌尿系疾病，尿液还可以反映全身各系统疾病状态，例如脑部疾病、心脏疾病、代谢性疾病。尿液中包涵丰富的信息，对泌尿系统，乃至整个机体系统疾病标记物的研究都具有重要的指导意义。
发明内容
本申请的技术方案基于这样的发现：在尿液中某些蛋白的表达水平和受试者肿瘤骨转移风险和/或是否存在肿瘤骨转移之间具有统计学意义上的显著相关性。
在一些实施方案中，尿液中的蛋白选自：
心房利钠肽受体2、半乳糖凝集素3结合蛋白、铁调素、骨桥蛋白、生长分化因子15、羟基酸氧化酶2、NKG2-DII型整合膜蛋白、脂肪酸结合蛋白、补体成分C6、α1酸性糖蛋白、硫氧还蛋白、β-2-微球蛋白、尿调节素、CMRF35样分子1、肿瘤坏死因子受体超家族成员1b、超氧化物歧化酶[Cu-Zn]、组织蛋白酶S、谷胱甘肽过氧化物酶3、1,2-二羟基-3-酮-5-甲硫基戊烯双加氧酶、细胞表面糖蛋白MUC18、纤维蛋白原α链、2-氧代戊二酸脱氢酶复合物的二氢脂酰赖氨酸残基琥珀酰基转移酶组分、溶酶体α-葡糖苷酶、棕榈酰蛋白硫酯酶1、前胶原赖氨酸-2-氧戊二酸5-双加氧酶3；
任选，HLA I类组织相容性抗原a-31α链、细胞色素c、前列腺素还原酶2、细胞间粘附分子-1、颗粒蛋白、含α/β水解酶结构域的蛋白14B、钠/葡萄糖协同转运蛋白1、糖基化依赖性细胞粘附分子1；
以及任选，金属硫蛋白-1、补体C4、粘附素细胞粘附分子1、铜转运蛋白ATOX1、6-磷酸葡糖酸内酯酶、7,8-二氢-8-氧鸟嘌呤三磷酸酶、凝血酶原、核糖核酸酶抑制剂、维生素D结合蛋白、凝固因子XII；
以及任选，V型质子ATP酶亚基B脑异构体、A激酶锚蛋白SPHKAP、Igλ-2链C区、L-乳酸脱氢酶C链、电压依赖性阴离子选择性通道蛋白1、卵泡抑素、双氢喋啶还原酶、低密度脂蛋白受体相关蛋白2、推定的磷脂酶B样2、γ-谷氨酰水解酶、载脂蛋白A-IV、血凝素、纤维连接蛋白、肽基甘氨酸α-酰胺化单氧酶、硫氧还蛋白、C反应蛋白、载脂蛋白C-I、26S蛋白酶调节亚单位7、软骨寡聚基质蛋白、二肽基肽酶2、脱氧核糖核酸酶1、凝溶胶蛋白、整膜蛋白2b、透明质酸酶-1、分泌皱褶相关蛋白4、活化素受体-1b型。
根据本申请的一些实施方案，提供了上述蛋白在预测受试者肿瘤骨转移风险和/或诊断受试者肿瘤骨转移中的用途。尤其是，上述蛋白在尿液中的表达水平在预测受试者肿瘤骨转移风险和/或诊断受试者肿瘤骨转移中的用途。
根据本申请的一些实施方案，提供了定量试剂在制备诊断装置中的用途，其中所述诊断装置用于预测受试者肿瘤骨转移风险和/或用于诊断受试者肿瘤骨转移；所述定量试剂是选自以下一种蛋白或其组合的定量试剂：
心房利钠肽受体2、半乳糖凝集素3结合蛋白、铁调素、骨桥蛋白、生长分化因子15、羟基酸氧化酶2、NKG2-DII型整合膜蛋白、脂肪酸结合蛋白、补体成分C6、α1酸性糖蛋白、硫氧还蛋白、β-2-微球蛋白、尿调节素、CMRF35样分子1、肿瘤坏死因子受体超家族成员1b、超氧化物歧化酶[Cu-Zn]、组织蛋白酶S、谷胱甘肽过氧化物酶3、1,2-二羟基-3-酮-5-甲硫基戊烯双加氧酶、细胞表面糖蛋白MUC18、纤维蛋白原α链、2-氧代戊二酸脱氢酶复合物的二氢脂酰赖氨酸残基琥珀酰基转移酶组分、溶酶体α-葡糖苷酶、棕榈酰蛋白硫酯酶1、前胶原赖氨酸-2-氧戊二酸5-双加氧酶3；
任选，HLA I类组织相容性抗原a-31α链、细胞色素c、前列腺素还原酶2、细胞间粘附分子-1、颗粒蛋白、含α/β水解酶结构域的蛋白14B、钠/葡萄糖协同转运蛋白1、糖基化依赖性细胞粘附分子1；
还任选，金属硫蛋白-1、补体C4、粘附素细胞粘附分子1、铜转运蛋白ATOX1、6-磷酸葡糖酸内酯酶、7,8-二氢-8-氧鸟嘌呤三磷酸酶、凝血酶原、核糖核酸酶抑制剂、维生素D结合蛋白、凝固因子XII；
还任选，V型质子ATP酶亚基B脑异构体、A激酶锚蛋白SPHKAP、Igλ-2链C区、L-乳酸脱氢酶C链、电压依赖性阴离子选择性通道蛋白1、卵泡抑素、双氢喋啶还原酶、低密度脂蛋白受体相关蛋白2、推定的磷脂酶B样2、γ-谷氨酰水解酶、载脂蛋白A-IV、血凝素、纤维连接蛋白、肽基甘氨酸α-酰胺化单氧酶、硫氧还蛋白、C反应蛋白、载脂蛋白C-I、26S蛋白酶调节亚单位7、软骨寡聚基质蛋白、二肽基肽酶2、脱氧核糖核酸酶1、凝溶胶蛋白、整膜蛋白2b、透明质酸酶-1、分泌皱褶相关蛋白4、活化素受体-1b型。
根据本申请的一些实施方案，尤其涉及上述蛋白在尿液中的表达水平在肿瘤骨转移风险早期诊断中的用途。
在一些实施方案中，定量试剂可以是已知的或未来的任何形式的定量试剂，只要它能够实现蛋白在尿液中表达水平的量化。例如，但不限于抗体或其抗原结合片段、质谱鉴定试剂。
在一些实施方案中，诊断装置体现为芯片或试剂盒的形式。
在本申请上下文中，对照是不具有肿瘤骨转移风险的群体或个体；或者不存在肿瘤骨转移的群体或个体。因此，在一些具体实施方案中，对照是例如健康群体或个体。在一些具体实施方案中，对照也可以指在这种群体或个体中蛋白的尿液表达水平。
在一些具体的实施方案中，相较于对照，选自以下任一种蛋白或其组合的表达水平升高，指示所述受试者肿瘤骨转移风险提高和/或诊断为肿瘤骨转移：心房利钠肽受体2、半乳糖凝集素-3结合蛋白、铁调素、骨桥蛋白、生长分化因子15、羟基酸氧化酶2、NKG2-DII型整合膜蛋白、脂肪酸结合蛋白、补体成分C6、α1酸性糖蛋白、线粒体硫氧还蛋白、β-2-微球蛋白、尿调节素、CMRF35样分子1、肿瘤坏死因子受体超家族成员1b、超氧化物歧化酶[Cu-Zn]、组织蛋白酶S、谷胱甘肽过氧化物酶3、1,2-二羟基-3-酮-5-甲硫基戊烯双加氧酶、细胞表面糖蛋白MUC18、纤维蛋白原α链、线粒体2-氧代戊二酸脱氢酶复合物的二氢脂酰赖氨酸残基琥珀酰基转移酶组分、溶酶体α-葡糖苷酶、HLA I类组织相容性抗原a-31α链、体细胞细胞色素c、前列腺素还原酶2、细胞间粘附分子-1、颗粒蛋白、含α/β水解酶结构域的蛋白质14B、金属硫蛋白-1、补体C4、粘附素细胞粘附分子1、铜转运蛋白ATOX1、6-磷酸葡糖酸内酯酶、7,8-二氢-8-氧鸟嘌呤三磷酸酶、凝血酶原、核糖核酸酶抑制剂、V型质子ATPase亚基B脑异构体、A激酶锚蛋白SPHKAP、Igλ-2链C区、L-乳酸脱氢酶C链、电压依赖性阴离子选择性通道蛋白1、卵泡抑素、双氢喋啶还原酶。
在另一些具体的实施方案中，相较于对照，选自以下任一种蛋白或其组合的表达水平降低，指示所述受试者肿瘤骨转移风险提高和/或诊断为肿瘤骨转移：
棕榈酰蛋白硫酯酶1、前胶原赖氨酸2-氧戊二酸5-双加氧酶3、钠/葡萄糖协同转运蛋白1、糖基化依赖性细胞粘附分子1、维生素D结合蛋白、凝固因子XII、低密度脂蛋白受体相关蛋白2、推定的磷脂酶B样2、γ-谷氨酰水解酶、载脂蛋白A-IV、血凝素、纤维连接蛋白、肽基甘氨酸α-酰胺化单氧酶、硫氧还蛋白、C反应蛋白、载脂蛋白C-I、26S蛋白酶调节亚单位7、软骨寡聚基质蛋白、二肽基肽酶2、脱氧核糖核酸酶1、凝溶胶蛋白、整膜蛋白2b、透明质酸酶-1、分泌皱褶相关蛋白4、活化素受体-1b型。
根据一些实施方案中，提供用于实施本申请方法或用途的诊断装置，其包含针对以下蛋白的定量试剂：
心房利钠肽受体2、半乳糖凝集素3结合蛋白、铁调素、骨桥蛋白、生长分化因子15、羟基酸氧化酶2、NKG2-DII型整合膜蛋白、脂肪酸结合蛋白、补体成分C6、α1酸性糖蛋白、硫氧还蛋白、β-2-微球蛋白、尿调节素、CMRF35样分子1、肿瘤坏死因子受体超家族成员1b、超氧化物歧化酶[Cu-Zn]、组织蛋白酶S、谷胱甘肽过氧化物酶3、1,2-二羟基-3-酮-5-甲硫基戊烯双加氧酶、细胞表面糖蛋白MUC18、纤维蛋白原α链、2-氧代戊二酸脱氢酶复合物的二氢脂酰赖氨酸残基琥珀酰基转移酶组分、溶酶体α-葡糖苷酶、棕榈酰蛋白硫酯酶1、前胶原赖氨酸-2-氧戊二酸5-双加氧酶3；
任选，HLA I类组织相容性抗原a-31α链、细胞色素c、前列腺素还原酶2、细胞间粘附分子-1、颗粒蛋白、含α/β水解酶结构域的蛋白14B、钠/葡萄糖协同转运蛋白1、糖基化依赖性细胞粘附分子1；
还任选，金属硫蛋白-1、补体C4、粘附素细胞粘附分子1、铜转运蛋白ATOX1、6-磷酸葡糖酸内酯酶、7,8-二氢-8-氧鸟嘌呤三磷酸酶、凝血酶原、核糖核酸酶抑制剂、维生素D结合蛋白、凝固因子XII；
还任选，V型质子ATPase亚基B脑异构体、A激酶锚蛋白SPHKAP、Igλ-2链C区、L-乳酸脱氢酶C链、电压依赖性阴离子选择性通道蛋白1、卵泡抑素、双氢喋啶还原酶低密度脂蛋白受体相关蛋白2、推定的磷脂酶B样2、γ-谷氨酰水解酶、载脂蛋白A-IV、血凝素、纤维连接蛋白、肽基甘氨酸α-酰胺化单氧酶、硫氧还蛋白、C反应蛋白、载脂蛋白C-I、26S蛋白酶调节亚单位7、软骨寡聚基质蛋白、二肽基肽酶2、脱氧核糖核酸酶1、凝溶胶蛋白、整膜蛋白2b、透明质酸酶-1、分泌皱褶相关蛋白4、活化素受体-1b型。
附图说明
图1显示对照大鼠与walker-256细胞种植到胫骨的大鼠体重变化。
图2A至图2C显示大鼠胫骨CT变化。依此为对照组(图2A)、D5(图2B)、D10(图2C)的CT结果。
具体实施方式
实施例1.公认骨转移模型的建立
本实施例中使用的Walker-256细胞是在怀孕大鼠(Rattus norvegicus)的乳腺中发现，它被视为一种癌肉瘤(参见McEuen,C.S等人(1933)Br.J.Exp.Pathol.14,384–391和Simpkins,H等人(1991)Cancer Res.51,1334–1338)。
Walker-256是实验研究中使用最广泛的可移植性肿瘤。该细胞在大鼠骨骼植入的位置能造成明显的骨吸收和骨破坏，与乳腺癌患者骨转移的表型一致，是本领域公认的骨转移模型(参见Shih,L.Y等人(2004)J.Orthop.Res.22,1161–1167)。在本实施例中，本发明人将walker-256细胞种植到大鼠的左腿胫骨，模拟肿瘤骨转移。在肿瘤生长前、初期、末期收取尿液进行分析。
1.材料与试剂
1)仪器：
Orbitrap Fusion Lumos Tribird质谱仪购自Thermo Scientific公司；大鼠代谢笼：购自北京佳源兴业科技有限公司。
2)主要试剂：
色谱质谱级水、乙腈、甲酸和甲醇从Fisher公司购买；碘乙酰铵(IAA)、碳酸氢铵、尿素、二硫苏糖醇(DTT)从Sigma公司购买；黄金胰酶从Promega公司购买。Walker-256细胞从国家实验细胞资源共享平台购买。
3)动物：
雄性Sprague-Dawley大鼠(体重180-200g)、SD幼鼠(60-80g)购自维通利华公司，在标准饲养环境中饲养(室温22±1℃，湿度65-70％)。
2.动物模型的建立和尿液样本收集
实验前对SD大鼠进行称重，将只大鼠随机分为两组。
实验组：大鼠禁食24小时后使用2％的戊巴比妥钠麻醉，用10ml注射器针头在大鼠左腿胫骨隆起处穿孔，用10μl微量进样器将5μl(2×104个/mL)的walker-256细胞注入骨髓腔内。注入后停顿30秒到1分钟，缓慢退针，使细胞充分进入骨髓腔，减少溢出。使用水门汀封堵针孔，缝合伤口处皮肤。
对照组：与实验组类似处理，骨髓腔内注射10μl生理盐水，而不是细胞溶液。取尿时，测量造模后的实验组大鼠和对照组大鼠的体重。
实施例2.尿液的样本收集和蛋白鉴定
1.尿液样本收集：
造模后，第3、5、7、10、13天将实验组与对照组大鼠置于代谢笼中收集尿液。并取3只大鼠，使用CT观察大鼠胫骨病变情况。
2.乙醇沉淀法提取尿液中蛋白
乙醇沉淀尿蛋白方法参照Sun W等方法(参见Sun W等人Proteomics,2005,5(18):4994–5001)。尿液离心：3000g/min、4℃、30min，取上清；12000g/min、4℃离心30min。取上清，加入3-5倍预冷的乙醇，4℃沉淀过夜。12000g/min、4℃离心30min，留沉淀室温风干。加入裂解液约500μl吹打溶解沉淀，超声3min。12000g/min、4℃离心30min，留取上清，Bradford法测蛋白浓度-80℃保存备用。
3.尿蛋白胰酶酶解
每个样本取100μg尿蛋白，按Wisniewski等方法酶解(参见Wisniewski JR等人Universal sample preparation method for proteome analysis.Nat Methods,2009,6:359-363)。加入DTT终浓度4.5mM、37℃、60min还原；加入IAA终浓度10mM、室温避光、30min烷基化；按照质量比例(尿液蛋白：胰酶＝50：1)加入胰酶，37℃、水浴、16小时。酶解后回收肽段用Oasis HLB小柱除盐，并冻干成粉末，于-20℃存储备用。
4.高效液相串联质谱法(LC-MS/MS)分析
将第3步所的肽段溶于0.1％甲酸，每个样品取900ng肽段进行色谱分离(ThermoEASY-nLC 1200)：
洗脱时间90min，色谱柱流速0.3μL/min，洗脱梯度为4％-28％流动相B(0.1％甲酸+79.9％乙腈+20％水)；反相柱(C18，长15cm，内径50μm)
洗脱下的多肽用Orbitrap Fusion Lumos质谱仪进行鉴定。喷雾电压2.1kV，离子传输管温度320℃。一级全扫描：350-1550m/z，120 000分辨率。二级扫描为数据依赖采集模式，循环时间3s，最高速度模式。其它参数：HCD碎裂方式，30％碎裂能量，30 000分辨率，扫描起始110m/z。每个样品进行2次技术重复鉴定。
5.数据库检索和数据分析
应用Mascot软件(2.4.0)进行数据库检索，方法参照(Perkins DN等Electrophoresis,1999,20)。具体如下：
-将质谱仪采集到的raw文件，进行二级谱谱峰提取，转换成MGF格式文件。
-将所有MGF格式文件导入到Mascot软件。
-设置搜库参数：所用数据库为Swissprot_database(数据截止至2017年6月3日)。
-其他检索条件为：物种设为大鼠；胰酶酶切，最多允许2个胰酶漏切位点；固定修饰为半胱氨酸的脲基甲基化，可变修饰为N末端乙酰化修饰和氧化修饰；母离子质量精确度为0.02Da，子离子质量精确度为10ppm。
-搜库后筛选条件：肽段水平FDR<1％，蛋白水平FDR<1％，ion score>30。最后将搜库结果以dat格式导出，以备后续定量分析。
将Orbitrap Fusion Lumos采集的数据(raw格式文件)直接导入用ProgenesisLC-MS软件(版本：4.1)。软件自动选择最合适的数据作为参照，对其它质谱数据进行保留时间的校正。
选择带2+、3+、4+电荷数的母离子进行后续定量。将所有普峰数据导出mgf格式文件，用Mascot软件(版本2.4.0)进行搜库。检索完成后，将p值设为0.01，Mascot打分设为30，导出xml格式文件，并导入progenesis。以鉴定到多肽的一级谱峰强度为基础进行定量分析。
基于二级谱谱图数进行定量：运用软件Scaffold软件(version 4.0.1,ProteomeSoftware Inc.,Portland,OR,USA)MASCOT 2.4.0软件搜库结果进行筛选整合，及谱图数定量。将Mascot软件搜库后导出的dat文件，逐一导入到Scaffold软件中。选择定量方法为：二级谱谱图数定量。数据库选择：与Mascot软件搜库应用同样的数据库，Swissprot_database(数据截止至2017年6月3日)。检索条件：肽段鉴定条件为可信度≥90.0％，FDR≤0.1％，蛋白鉴定条件为可信度≥95.0％，并且每个蛋白包括至少2个鉴定到的肽段(参见Elias JE等人Nat Methods,2007,4:207-214)。
6.实验结果
1)大鼠临床表现与体重变化
正常对照组大鼠活跃，毛发光泽，大小便正常。Walker-256细胞种植胫骨组大鼠于建模第7天出现活动减少、饮食减少。骨转移组大鼠体重，从第10天开始明显低于对照组(图1)。
2)胫骨CT分析结果
在第5、7、10、14天分别取实验组3只大鼠，对照组1只大鼠。使用SIEMENS Inveon临床前(小动物)PET-CT，对大鼠左腿胫骨拍摄CT，观察骨密度及破损情况的进展。
对照组：未发现放射学变化(图2A)。
实验组第5天：肿瘤细胞在针孔附近聚集生长，未见明显骨破坏(图2B)。
实验组第10天：肿瘤细胞侵袭性生长，骨密度明显下降，骨破坏明显(图2C)。
3)尿液差异表达蛋白筛选
来源于大鼠5个时期(对照组n＝6，实验组第3、5、7、13天各n＝6)的30个尿液样本。首先，经过蛋白组学分析鉴定。筛选蛋白条件为FDR小于1％的水平，并且每个蛋白至少有1条特异的肽段，共鉴定到954个蛋白。差异表达的蛋白筛选标准：蛋白丰度变化倍数在2倍以上，p值小于0.05，实验组每一个归一化丰度(Normalized abundance)均大于或小于对照组每一个归一化丰度。同时使用Uniprot数据库将大鼠蛋白转化为相应的人同源蛋白。在本实施例所采用的鉴定方法中，每一个鉴定出的差异表达蛋白均与实验分组(对照vs骨转移组)具有独立的统计学意义，这指示这每一个蛋白标记物可以作为独立的预测因子，也可以组合使用。
第3天实验组与对照组相比，一共鉴定到25个差异表达的蛋白，有23个蛋白表达上调，2个蛋白表达下调。
第5天实验组与对照组相比，一共鉴定到13个差异表达的蛋白，有10个蛋白表达上调，3个蛋白表达下调。
第7天实验组与对照组相比，一共鉴定到21个差异表达的蛋白，有18个蛋白表达上调，3个蛋白表达下调。
第13天实验组与对照组相比，一共鉴定到27个差异表达的蛋白，有7个蛋白表达上调，20个蛋白表达下调。
表1.肿瘤骨转移尿液蛋白标记物
Claims (10)
1.定量试剂在制备诊断装置中的用途，其中：
所述诊断装置用于预测受试者肿瘤骨转移风险和/或用于诊断受试者肿瘤骨转移；
所述定量试剂是选自以下一种蛋白或其组合的定量试剂：
心房利钠肽受体2、半乳糖凝集素3结合蛋白、铁调素、骨桥蛋白、生长分化因子15、羟基酸氧化酶2、NKG2-DII型整合膜蛋白、脂肪酸结合蛋白、补体成分C6、α1酸性糖蛋白、硫氧还蛋白、β-2-微球蛋白、尿调节素、CMRF35样分子1、肿瘤坏死因子受体超家族成员1b、超氧化物歧化酶[Cu-Zn]、组织蛋白酶S、谷胱甘肽过氧化物酶3、1,2-二羟基-3-酮-5-甲硫基戊烯双加氧酶、细胞表面糖蛋白MUC18、纤维蛋白原α链、2-氧代戊二酸脱氢酶复合物的二氢脂酰赖氨酸残基琥珀酰基转移酶组分、溶酶体α-葡糖苷酶、棕榈酰蛋白硫酯酶1、前胶原赖氨酸-2-氧戊二酸5-双加氧酶3；
所述定量试剂确定所述蛋白在尿液中的表达水平；
优选，所述诊断是早期诊断。
2.根据权利要求1所述的用途，所述蛋白还选自以下一种或其组合：
HLA I类组织相容性抗原a-31α链、细胞色素c、前列腺素还原酶2、细胞间粘附分子-1、颗粒蛋白、含α/β水解酶结构域的蛋白14B、钠/葡萄糖协同转运蛋白1、糖基化依赖性细胞粘附分子1。
3.根据权利要求1或2所述的用途，所述蛋白还选自以下一种或其组合：
金属硫蛋白-1、补体C4、粘附素细胞粘附分子1、铜转运蛋白ATOX1、6-磷酸葡糖酸内酯酶、7,8-二氢-8-氧鸟嘌呤三磷酸酶、凝血酶原、核糖核酸酶抑制剂、维生素D结合蛋白、凝固因子XII。
4.根据权利要求1至3任一项所述的用途，所述蛋白还选自以下一种或其组合：
V型质子ATP酶亚基B脑异构体、A激酶锚蛋白SPHKAP、Igλ-2链C区、L-乳酸脱氢酶C链、电压依赖性阴离子选择性通道蛋白1、卵泡抑素、双氢喋啶还原酶、低密度脂蛋白受体相关蛋白2、推定的磷脂酶B样2、γ-谷氨酰水解酶、载脂蛋白A-IV、血凝素、纤维连接蛋白、肽基甘氨酸α-酰胺化单氧酶、硫氧还蛋白、C反应蛋白、载脂蛋白C-I、26S蛋白酶调节亚单位7、软骨寡聚基质蛋白、二肽基肽酶2、脱氧核糖核酸酶1、凝溶胶蛋白、整膜蛋白2b、透明质酸酶-1、分泌皱褶相关蛋白4、活化素受体-1b型。
5.根据权利要求1至4任一项所述的用途，其中所述定量试剂选自：抗体或其抗原结合片段、质谱鉴定试剂。
6.根据权利要求1所述的用途，其中所述诊断装置是芯片或试剂盒。
7.根据权利要求1至4任一项所述的用途，其中：
相较于对照，选自以下任一种蛋白或其组合的表达水平升高，指示所述受试者肿瘤骨转移风险提高和/或诊断为肿瘤骨转移：
心房利钠肽受体2、半乳糖凝集素-3结合蛋白、铁调素、骨桥蛋白、生长分化因子15、羟基酸氧化酶2、NKG2-DII型整合膜蛋白、脂肪酸结合蛋白、补体成分C6、α1酸性糖蛋白、线粒体硫氧还蛋白、β-2-微球蛋白、尿调节素、CMRF35样分子1、肿瘤坏死因子受体超家族成员1b、超氧化物歧化酶[Cu-Zn]、组织蛋白酶S、谷胱甘肽过氧化物酶3、1,2-二羟基-3-酮-5-甲硫基戊烯双加氧酶、细胞表面糖蛋白MUC18、纤维蛋白原α链、线粒体2-氧代戊二酸脱氢酶复合物的二氢脂酰赖氨酸残基琥珀酰基转移酶组分、溶酶体α-葡糖苷酶、HLA I类组织相容性抗原a-31α链、体细胞细胞色素c、前列腺素还原酶2、细胞间粘附分子-1、颗粒蛋白、含α/β水解酶结构域的蛋白质14B、金属硫蛋白-1、补体C4、粘附素细胞粘附分子1、铜转运蛋白ATOX1、6-磷酸葡糖酸内酯酶、7,8-二氢-8-氧鸟嘌呤三磷酸酶、凝血酶原、核糖核酸酶抑制剂、V型质子ATPase亚基B脑异构体、A激酶锚蛋白SPHKAP、Igλ-2链C区、L-乳酸脱氢酶C链、电压依赖性阴离子选择性通道蛋白1、卵泡抑素、双氢喋啶还原酶；
或者，
相较于对照，选自以下任一种蛋白或其组合的表达水平降低，指示所述受试者肿瘤骨转移风险提高和/或诊断为肿瘤骨转移：
棕榈酰蛋白硫酯酶1、前胶原赖氨酸2-氧戊二酸5-双加氧酶3、钠/葡萄糖协同转运蛋白1、糖基化依赖性细胞粘附分子1、维生素D结合蛋白、凝固因子XII、低密度脂蛋白受体相关蛋白2、推定的磷脂酶B样2、γ-谷氨酰水解酶、载脂蛋白A-IV、血凝素、纤维连接蛋白、肽基甘氨酸α-酰胺化单氧酶、硫氧还蛋白、C反应蛋白、载脂蛋白C-I、26S蛋白酶调节亚单位7、软骨寡聚基质蛋白、二肽基肽酶2、脱氧核糖核酸酶1、凝溶胶蛋白、整膜蛋白2b、透明质酸酶-1、分泌皱褶相关蛋白4、活化素受体-1b型。
8.一种诊断装置，其包含针对以下蛋白的组合的定量试剂：
心房利钠肽受体2、半乳糖凝集素3结合蛋白、铁调素、骨桥蛋白、生长分化因子15、羟基酸氧化酶2、NKG2-DII型整合膜蛋白、脂肪酸结合蛋白、补体成分C6、α1酸性糖蛋白、硫氧还蛋白、β-2-微球蛋白、尿调节素、CMRF35样分子1、肿瘤坏死因子受体超家族成员1b、超氧化物歧化酶[Cu-Zn]、组织蛋白酶S、谷胱甘肽过氧化物酶3、1,2-二羟基-3-酮-5-甲硫基戊烯双加氧酶、细胞表面糖蛋白MUC18、纤维蛋白原α链、2-氧代戊二酸脱氢酶复合物的二氢脂酰赖氨酸残基琥珀酰基转移酶组分、溶酶体α-葡糖苷酶、棕榈酰蛋白硫酯酶1、前胶原赖氨酸-2-氧戊二酸5-双加氧酶3。
9.根据权利要求8所述的诊断装置，还包含针对选自以下一种蛋白或其组合的定量试剂：
HLA I类组织相容性抗原a-31α链、细胞色素c、前列腺素还原酶2、细胞间粘附分子-1、颗粒蛋白、含α/β水解酶结构域的蛋白14B、钠/葡萄糖协同转运蛋白1、糖基化依赖性细胞粘附分子1；
任选，所述诊断装置还包含针对选自以下一种蛋白或其组合的定量试剂：金属硫蛋白-1、补体C4、粘附素细胞粘附分子1、铜转运蛋白ATOX1、6-磷酸葡糖酸内酯酶、7,8-二氢-8-氧鸟嘌呤三磷酸酶、凝血酶原、核糖核酸酶抑制剂、维生素D结合蛋白、凝固因子XII；
任选，所述诊断装置还包含针对选自以下一种蛋白或其组合的定量试剂：V型质子ATPase亚基B脑异构体、A激酶锚蛋白SPHKAP、Igλ-2链C区、L-乳酸脱氢酶C链、电压依赖性阴离子选择性通道蛋白1、卵泡抑素、双氢喋啶还原酶低密度脂蛋白受体相关蛋白2、推定的磷脂酶B样2、γ-谷氨酰水解酶、载脂蛋白A-IV、血凝素、纤维连接蛋白、肽基甘氨酸α-酰胺化单氧酶、硫氧还蛋白、C反应蛋白、载脂蛋白C-I、26S蛋白酶调节亚单位7、软骨寡聚基质蛋白、二肽基肽酶2、脱氧核糖核酸酶1、凝溶胶蛋白、整膜蛋白2b、透明质酸酶-1、分泌皱褶相关蛋白4、活化素受体-1b型。
10.根据权利要求8所述的诊断装置，其中：
所述定量试剂选自：抗体或其抗原结合片段、质谱鉴定试剂；
所述诊断装置是芯片或试剂盒。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910289068.9A CN111812309A (zh) | 2019-04-11 | 2019-04-11 | 肿瘤骨转移的尿液蛋白标记物及其用途 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910289068.9A CN111812309A (zh) | 2019-04-11 | 2019-04-11 | 肿瘤骨转移的尿液蛋白标记物及其用途 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111812309A true CN111812309A (zh) | 2020-10-23 |
Family
ID=72843809
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910289068.9A Pending CN111812309A (zh) | 2019-04-11 | 2019-04-11 | 肿瘤骨转移的尿液蛋白标记物及其用途 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111812309A (zh) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111308095A (zh) * | 2020-03-04 | 2020-06-19 | 北京师范大学 | 用于诊断前列腺癌的尿液蛋白标记物 |
| CN112964879A (zh) * | 2021-02-03 | 2021-06-15 | 北京师范大学 | 与动脉粥样硬化相关的尿液蛋白标志物、其获得方法及其用途 |
-
2019
- 2019-04-11 CN CN201910289068.9A patent/CN111812309A/zh active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111308095A (zh) * | 2020-03-04 | 2020-06-19 | 北京师范大学 | 用于诊断前列腺癌的尿液蛋白标记物 |
| CN112964879A (zh) * | 2021-02-03 | 2021-06-15 | 北京师范大学 | 与动脉粥样硬化相关的尿液蛋白标志物、其获得方法及其用途 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Vitório et al. | Metabolic landscape of oral squamous cell carcinoma | |
| Sigdel et al. | Shotgun proteomics identifies proteins specific for acute renal transplant rejection | |
| US20170168057A1 (en) | Srm assays to chemotherapy targets | |
| WO2014144499A1 (en) | Method for replacing biomarkers of protein kinetics from tissue samples by biomarkers of protein kinetics from body fluids after isotopic labeling in vivo | |
| CN111812309A (zh) | 肿瘤骨转移的尿液蛋白标记物及其用途 | |
| WO2017066780A1 (en) | Sulfatide analysis for newborn screening and diagnosis of metachromatic leukodystrophy | |
| CN110554134A (zh) | 临床血液样本分析方法及应用 | |
| US20130289142A1 (en) | Her3 protein srm/mrm assay | |
| CN107870245A (zh) | 基于唾液蛋白质组学的慢性肾炎生物标志物检测的方法 | |
| US20160349265A1 (en) | Insulin Receptor Substrate 1 (IRS1) Protein SRM/MRM Assay | |
| CN110268263A (zh) | 人体皮肤老化的新生物标志物 | |
| EP3859339A1 (en) | Mild-cognitive-impairment evaluation method, calculation method, evaluation device, calculation device, evaluation program, calculation program, recording medium, evaluation system, and terminal device | |
| Yang et al. | Recent advances in protein profiling of tissues and tissue fluids | |
| CN108918868A (zh) | 一种检测骨关节炎的标记物及其应用 | |
| Golea-Secara et al. | Urinary proteins detected using modern proteomics intervene in early type 2 diabetic kidney disease–a pilot study | |
| CN116754772A (zh) | 老年痴呆早期诊断外周血蛋白标志物、应用及辅助诊断系统 | |
| JP2011512149A (ja) | ガンの検出の方法および手法 | |
| JP2009128240A (ja) | タンパク質試料中のメチオニン残基の酸化状態を解析する方法 | |
| CN112924692B (zh) | 一种基于多肽定量测定的糖尿病诊断试剂盒及其方法 | |
| CN109975546B (zh) | 早期乳腺癌肺转移相关的尿蛋白标志物的筛选及其用途 | |
| JP6809704B2 (ja) | 酸化ストレスマーカー及びその使用 | |
| Zhang et al. | Serum proteomic analysis of differentially expressed proteins and pathways involved in the mechanism of endemic osteoarthritis | |
| JP2009210355A (ja) | 肺腺癌予後診断マーカー | |
| CN112924689A (zh) | 基于多肽组合标志物定量测定的糖尿病诊断试剂盒及其方法 | |
| US20130072581A1 (en) | Secreted Protein Acidic and Rich in Cysteine (SPARC) Protein SRM Assay |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20201023 |
|
| WD01 | Invention patent application deemed withdrawn after publication |