CN117210563A - 一种乳腺癌诊断和/或预后判断用mRNA标志物及应用 - Google Patents
一种乳腺癌诊断和/或预后判断用mRNA标志物及应用 Download PDFInfo
- Publication number
- CN117210563A CN117210563A CN202311062278.7A CN202311062278A CN117210563A CN 117210563 A CN117210563 A CN 117210563A CN 202311062278 A CN202311062278 A CN 202311062278A CN 117210563 A CN117210563 A CN 117210563A
- Authority
- CN
- China
- Prior art keywords
- breast cancer
- prss22
- marker
- prognosis
- expression
- 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
- 206010006187 Breast cancer Diseases 0.000 title claims abstract description 86
- 208000026310 Breast neoplasm Diseases 0.000 title claims abstract description 85
- 238000004393 prognosis Methods 0.000 title claims abstract description 22
- 239000003550 marker Substances 0.000 title claims abstract description 21
- 108020004999 messenger RNA Proteins 0.000 title claims abstract description 14
- 238000003745 diagnosis Methods 0.000 title claims abstract description 11
- 102100022046 Brain-specific serine protease 4 Human genes 0.000 claims abstract description 79
- 101000896891 Homo sapiens Brain-specific serine protease 4 Proteins 0.000 claims abstract description 78
- 239000000523 sample Substances 0.000 claims description 13
- 239000003153 chemical reaction reagent Substances 0.000 claims description 5
- 238000012360 testing method Methods 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 3
- 239000012472 biological sample Substances 0.000 claims description 2
- 230000002934 lysing effect Effects 0.000 claims description 2
- 210000000056 organ Anatomy 0.000 claims description 2
- 230000014509 gene expression Effects 0.000 abstract description 48
- 206010027476 Metastases Diseases 0.000 abstract description 8
- 230000009401 metastasis Effects 0.000 abstract description 7
- 238000010837 poor prognosis Methods 0.000 abstract description 6
- 201000010099 disease Diseases 0.000 abstract description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 5
- 238000001514 detection method Methods 0.000 abstract description 3
- 238000003759 clinical diagnosis Methods 0.000 abstract description 2
- 206010059866 Drug resistance Diseases 0.000 abstract 1
- 238000013399 early diagnosis Methods 0.000 abstract 1
- 210000001519 tissue Anatomy 0.000 description 46
- 210000004027 cell Anatomy 0.000 description 18
- 208000007433 Lymphatic Metastasis Diseases 0.000 description 15
- 230000004083 survival effect Effects 0.000 description 15
- 206010028980 Neoplasm Diseases 0.000 description 11
- 210000000481 breast Anatomy 0.000 description 11
- 230000008595 infiltration Effects 0.000 description 9
- 238000001764 infiltration Methods 0.000 description 9
- 230000005012 migration Effects 0.000 description 8
- 238000013508 migration Methods 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 6
- 230000002018 overexpression Effects 0.000 description 6
- 240000005528 Arctium lappa Species 0.000 description 5
- 201000011510 cancer Diseases 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 238000011529 RT qPCR Methods 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 239000013598 vector Substances 0.000 description 4
- 206010061535 Ovarian neoplasm Diseases 0.000 description 3
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 3
- 238000011282 treatment Methods 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 206010014733 Endometrial cancer Diseases 0.000 description 2
- 206010014759 Endometrial neoplasm Diseases 0.000 description 2
- 206010018338 Glioma Diseases 0.000 description 2
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 2
- 206010033128 Ovarian cancer Diseases 0.000 description 2
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 2
- 238000003559 RNA-seq method Methods 0.000 description 2
- 108010022999 Serine Proteases Proteins 0.000 description 2
- 102000012479 Serine Proteases Human genes 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000013211 curve analysis Methods 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 238000000556 factor analysis Methods 0.000 description 2
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 2
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 201000002528 pancreatic cancer Diseases 0.000 description 2
- 208000008443 pancreatic carcinoma Diseases 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000001356 surgical procedure Methods 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 101710135671 Brain-specific serine protease 4 Proteins 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 208000032612 Glial tumor Diseases 0.000 description 1
- 239000012097 Lipofectamine 2000 Substances 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 206010027459 Metastases to lymph nodes Diseases 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000015634 Rectal Neoplasms Diseases 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000012292 cell migration Effects 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 208000006990 cholangiocarcinoma Diseases 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000010219 correlation analysis Methods 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000013020 embryo development Effects 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 230000020764 fibrinolysis Effects 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- 238000011223 gene expression profiling Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 108010082117 matrigel Proteins 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000001617 migratory effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 206010038038 rectal cancer Diseases 0.000 description 1
- 201000001275 rectum cancer Diseases 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000004017 serum-free culture medium Substances 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 239000012096 transfection reagent Substances 0.000 description 1
- 206010044412 transitional cell carcinoma Diseases 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 208000023747 urothelial carcinoma Diseases 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
-
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
- G01N33/57415—Specifically defined cancers of breast
-
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/118—Prognosis of disease development
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
- G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
- G01N2333/95—Proteinases, i.e. endopeptidases (3.4.21-3.4.99)
- G01N2333/964—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue
- G01N2333/96425—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals
- G01N2333/96427—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general
- G01N2333/9643—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general with EC number
- G01N2333/96433—Serine endopeptidases (3.4.21)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Cell Biology (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Food Science & Technology (AREA)
- General Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Hospice & Palliative Care (AREA)
- Oncology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Biophysics (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明涉及一种乳腺癌诊断和/或预后判断用mRNA标志物及应用,属于分子生物学和疾病诊断技术领域。乳腺癌对女性健康构成了重要威胁,该疾病耐药、转移和复发的频率较高,晚期乳腺癌患者预后状况不佳,提供一种早期诊断标志物对避免乳腺癌的不良预后具有重要意义。本发明提供了PRSS22作为乳腺癌诊断和/或预后判断用标志物的应用,该标志物在病灶组织和癌旁组织中表达量具有明显的差异,并且与乳腺癌的不良预后相关,对于乳腺癌的临床诊断及不良预后防治具有重要的检测价值。
Description
技术领域
本公开属于分子生物学和疾病诊断技术领域,具体涉及一种乳腺癌诊断和/或预后判断用mRNA标志物及应用。
背景技术
乳腺癌是世界上常见的恶性肿瘤之一,已成为女性癌症死亡的主要原因。乳腺癌转移是导致患者临床治疗失败的重要原因,严重威胁着患者的生命。寻找乳腺癌诊断和预后的标志物有利于早期发现疾病,并筛选可能预后不良的高危人群,制定疾病的临床诊疗策略,为乳腺癌的治疗提供新的思路。
PRSS22全称称为类胰蛋白酶ε和丝氨酸蛋白酶S1家族成员22,而丝氨酸蛋白酶最初被发现在胚胎发育、免疫、凝血、受精、纤维蛋白溶解、激素激活、食物消化及其他生物过程中起重要作用。近年来,其在肿瘤中的表达及作用逐渐引起了研究者的重视。虽然PRSS22已被证明在多种恶性肿瘤如胶质瘤、胰腺癌、前列腺癌、卵巢癌中存在表达,但其在乳腺癌中的表达情况及预后价值尚不清楚。
发明内容
针对上述研究背景,本领域内虽然PRSS22已被证明在多种恶性肿瘤如胶质瘤、胰腺癌、前列腺癌、卵巢癌中存在表达,但其在乳腺癌中的表达情况及预后价值尚不清楚。发明人对PRSS22与乳腺癌之间的作用关系进行了研究,研究结果表明PRSS22的表达含量与乳腺癌患者的预后情况存在相关性,有望作为乳腺癌预后判断及治疗的新型标志物。
为了实现上述目的,本发明的技术方案为:
本发明的第一方面,一种乳腺癌诊断和/或预后判断用mRNA标志物,所述mRNA标志物为PRSS22。
所述标志物选自受试者乳腺癌组织。
本发明通过研究发现PRSS22在乳腺癌中表达上调,PRSS22高表达的患者较低表达的患者预后不良,PRSS22是独立预后因子。
本发明的第二方面,检测PRSS22的试剂组合在制备乳腺癌诊断和/或预后产品中的应用。
检测上述标志物的物质可以是采用诸如ELISA、胶体金试纸条、蛋白芯片等现有检测待测样本中相关蛋白及多肽表达情况的物质。
所述待测样本为人源样本;更具体的,所述待测样本为乳腺癌组织。
本发明的有益效果为:
上述技术方案提供一种乳腺癌诊断和/或预后判断用mRNA标志物及应用。上述技术方案通过研究发现,PRSS22在乳腺癌患者的乳腺组织中表达上调,PRSS22高表达的患者较低表达的患者预后不良,PRSS22是独立预后因子。PRSS22在乳腺癌组织较乳腺正常组织表达明显上调,ROC曲线显示其可以较好地区分癌组织与正常组织。与不伴有淋巴结转移的乳腺癌组织相比,PRSS22在伴有淋巴结转移的乳腺癌组织中表达较高,且PRSS22表达随淋巴结转移数目的增多逐渐升高;ROC曲线显示其可以较好地区分有无淋巴结转移的癌组织。使用PRSS22表达量的中位数作为临界值,将患者分为PRSS22低表达组与高表达组,发现其表达与年龄、组织学分级、肿瘤直径、ER、PR、HER2、Ki-67等其它临床病理参数无明显相关性。PRSS22过表达可显著促进乳腺癌细胞的迁移浸润能力。因此,PRSS22以作为乳腺癌潜在的诊断和/或预后判断用标记物,因此具有良好的实际应用之价值。
附图说明
构成本发明的一部分的说明书附图用来提供对本发明的进一步理解,本发明的示意性实施例及其说明用于解释本发明,并不构成对本发明的不当限定。
图1为本发明实施例中PRSS22 mRNA在乳腺癌组织及正常乳腺组织中的表达情况中GEPIA数据;
图2为本发明实施例中PRSS22 mRNA在乳腺癌组织及正常乳腺组织中的表达情况中UALCAN数据;
图3PRSS22 mRNA在乳腺癌组织及正常乳腺组织中的表达情况;
其中,A为PRSS22在乳腺癌组织较正常乳腺组织中表达上调;B为PRSS22区分乳腺癌组织与正常乳腺组织的ROC曲线图;C为PRSS22在伴有淋巴结转移的乳腺癌组织较不伴有乳腺癌组织中表达升高;随着淋巴结转移数目的增多,PRSS22表达逐渐升高;D为PRSS22区分有无淋巴结转移乳腺癌组织的ROC曲线图;
图4为本发明实施例中PRSS22与乳腺癌患者预后不良相关情况中Kaplan Meierplotter数据;
其中,A为乳腺癌患者PRSS22表达量高低与总生存时间关系图,B为乳腺癌患者PRSS22表达量高低与无远处转移生存时间关系图,C为乳腺癌患者PRSS22表达量高低与复发后生存时间关系图;
图5为本发明实施例中PRSS22与乳腺癌患者预后不良相关情况中GOBO数据;
其中,A为乳腺癌患者PRSS22表达量高低与总生存率关系图,B为多因素分析乳腺癌患者总生存率图,C为乳腺癌患者PRSS22表达量高低与无远处转移生存率关系图,D为多因素分析乳腺癌患者无远处转移生存率图;
图6为本发明实施例2中PRSS22过表达载体转染可显著上调PRSS22在乳腺癌细胞中的表达的数据;
图7为本发明实施例2中PRSS22促进乳腺癌细胞的迁移浸润能力的数据;
其中,A-B为乳腺癌细胞MDA-MB-231的迁移浸润图及统计柱状图,C-D为乳腺癌细胞MDA-MB-468的迁移浸润图及统计柱状图。
具体实施方式
应该指出,以下详细说明都是示例性的,旨在对本发明提供进一步的说明。除非另有指明,本文使用的所有技术和科学术语具有与本发明所属技术领域的普通技术人员通常理解的相同含义。
需要注意的是,这里所使用的术语仅是为了描述具体实施方式,而非意图限制根据本发明的示例性实施方式。如在这里所使用的,除非上下文另外明确指出,否则单数形式也意图包括复数形式,此外,还应当理解的是,当在本说明书中使用术语“包含”和/或“包括”时,其指明存在特征、步骤、操作、器件、组件和/或它们的组合。
本发明的一个典型具体实施方式中,提供一种乳腺癌诊断和/或预后判断用mRNA标志物,所述mRNA标志物为PRSS22。
本发明的一个或多个具体实施方式中,所述标志物选自受试者乳腺组织或乳腺癌组织。
本发明通过研究发现PRSS22在乳腺癌中表达上调,PRSS22高表达的患者较低表达的患者预后不良,PRSS22是独立预后因子。
本发明的第二个典型具体实施方式中,提供检测PRSS22的试剂组合在制备乳腺癌诊断和/或预后产品中的应用。
本发明的一个或多个具体实施方式中,所述产品包括试剂盒。
本发明的一个或多个具体实施方式中,检测样本为患者的临床生物样本,包括组织、器官中的一种或几种。
本发明的一个或多个具体实施方式中,所述检测样本为乳腺癌组织。
本发明的一个或多个具体实施方式中,所述试剂组合中至少包括如下试剂:
(1)引物;
(2)样本组织裂解试剂;
(3)PCR反应体系。
本发明的一个或多个具体实施方式中,所述引物包括PRSS22引物;所述PRSS22引物序列如下:
PRSS22-F:TCGACAGCCATCCTCAATGC(SEQ ID NO.1);
PRSS22-R:GTGGGTCCCATTCTTCTGGA(SEQ ID NO.2);
GAPDH-F:GCACCGTCAAGGCTGAGAAC(SEQ ID NO.3);
GAPDH-R:TGGTGAAGACGCCAGTGGA(SEQ ID NO.4)。
为了使得本领域技术人员能够更加清楚地了解本发明的技术方案,以下将结合具体的实施例详细说明本发明的技术方案。
实施例1
一种乳腺癌诊断和/或预后判断用mRNA标志物的确定
GEPIA(http://gepia.cancer-pku.cn/index.html)是一个基因表达谱分析的网站,它整合了基于TCGA等数据库的RNA测序信息,可进行肿瘤与正常组织的基因差异表达分析。
UALCAN(http://ualcan.path.uab.edu/)是一个基于TCGA基因表达数据分析的网站,其整合了来源于TCGA的31种肿瘤的RNA测序及临床信息。
Kaplan Meier plotter(http://kmplot.com/analysis/index.php?p=background)是一可做生存分析的网络数据库,GOBO数据库(http://co.bmc.lu.se/gobo/gsa.pl)基于Affymetrix U133A芯片平台的乳腺肿瘤芯片数据,可进行单个及多个基因的表达水平与预后的关联分析。
(1)使用GEPIA、UALCAN分析PRSS22在乳腺癌等多种肿瘤及正常组织中的表达情况。
(2)基于Kaplan Meier plotter、GOBO数据分析PRSS22在乳腺癌中的预后价值。
如图1所示,GEPIA数据显示PRSS22在乳腺癌、宫颈癌、卵巢癌、子宫内膜癌、直肠癌、结肠癌、胆管癌、胃癌、胰腺癌、子宫内膜癌、肺癌、膀胱尿路上皮癌等数十种人类常见恶性肿瘤如中表达水平均高于其对应正常组织。
如图2所示,UALCAN数据显示PRSS22表达在乳腺癌组织(n=1097)中明显高于正常乳腺组织(n=114,P<0.001)。
组织标本收集:于山东大学齐鲁医院收集56例乳腺癌新鲜组织及10例正常乳腺组织(2004年-2016年),组织来源于行手术治疗的乳腺癌患者。HE切片经高年资医师复核诊断,术前均未行放、化疗及其他针对肿瘤的治疗。患者均为女性,34-79岁,中位年龄52.5岁。组织储存于液氮中直至使用。
实时定量PCR:用Trizol提取总RNA,测定各组RNA的浓度。用TOYOBO反转录试剂盒ReverTra Ace qPCR RT Kit反转录成cDNA,以GAPDH作为内参,使用Roche Universal SYBRGreen Master进行实时定量PCR检测。
引物设计如下:
PRSS22-F:5’-TCGACAGCCATCCTCAATGC-3’(SEQ ID NO.1);
PRSS22-R:5’-GTGGGTCCCATTCTTCTGGA-3’(SEQ ID NO.2);
GAPDH-F:5’-GCACCGTCAAGGCTGAGAAC-3’(SEQ ID NO.3);
GAPDH-R:5’-TGGTGAAGACGCCAGTGGA-3’(SEQ ID NO.4)。
△CT=CT(PRSS22)-CT(GAPDH),根据相对表达量=2-△CT计算PRSS22 mRNA的表达。
如图3所示,PRSS22在乳腺癌组织较乳腺正常组织表达明显上调(图3A,P=0.0260)。ROC曲线分析显示,PRSS22可以较好地区分乳腺癌组织及正常组织(图3B,曲线下面积AUC=0.7232,P=0.02543),以0.006859为cutoff值,其敏感性、特异性分别为66.07%、80%。与不伴有淋巴结转移的乳腺癌组织相比,PRSS22在伴有淋巴结转移的乳腺癌组织中表达较高(图3C,P=
0.0284),且PRSS22表达随淋巴结转移数目的增多逐渐升高(图3C,P=0.0410)。ROC曲线分析显示,PRSS22可以较好地区分有无淋巴结转移组织(图3D,曲线下面积AUC=0.6723,P=0.02783),以0.005094为cutoff值,其敏感性、特异性分别为35.48%、100%。使用PRSS22表达量的中位数作为临界值,将患者分为PRSS22低表达组与高表达组,发现其表达与年龄、组织学分级、肿瘤直径、ER、PR、HER2、Ki-67等其它临床病理参数无明显相关性(表1)。
表1.PRSS22表达与患者临床病理参数的关系
如图4所示,Kaplan Meier plotter数据显示PRSS22高表达的乳腺癌患者的总生存时间(图4A,HR=1.29(1.03-1.61),P=0.024)、无远处转移生存时间(图4B,HR=1.35(1.09-1.66),P=0.0051)及复发后生存时间(图4C,HR=1.31(1.02-1.68),P=0.035)明显少于PRSS22低表达的患者。
如图5所示,GOBO数据显示PRSS22高表达的乳腺癌患者的总生存率(图5A,P=0.00158)及无远处转移生存率(图5C,P=0.00738)明显差于PRSS22低表达的患者。多因素分析(图5B)发现PRSS22、年龄、肿瘤直径、组织学分级及淋巴结转移是乳腺癌患者总生存的独立预后因子,其中PRSS22低表达提示预后较好(HR=0.66(0.5-0.89),P=0.006)。多因素分析(图5D)发现PRSS22、组织学分级及淋巴结转移是乳腺癌患者无远处转移生存的独立预后因子,其中PRSS22低表达提示预后较好(HR=0.66(0.49-0.88),P=0.005)。
实施例2
细胞培养及转染:细胞常规培养于含10%胎牛血清的L15或DMEM培养基中,在饱和湿度、5% CO2、37℃细胞培养箱中单层贴壁传代培养。使用Lipofectamine 2000转染试剂将PRSS22过表达载体(pcDNA3.1(+)-PRSS22)及对照载体(pcDNA3.1(+))转入乳腺癌细胞。
迁移浸润实验:细胞的迁移和浸润实验通过Transwell小室完成。迁移实验首先将转染后的乳腺癌细胞收集,用无血清培养基悬浮后,然后加入Transwell小室内,小室外再使用含血清的培养基孵育24h,经固定染色后显微镜下观察,计数迁移的细胞数。而浸润实验则提前在小室内铺好一层Matrigel matrix(基质胶),然后按照上述迁移实验的方法进行。
由于PRSS22在乳腺癌组织中的表达证实与淋巴结转移相关,本发明拟进一步探究其对乳腺癌细胞迁移浸润能力的影响。如图6所示,构建PRSS22过表达载体后,将PRSS22过表达载体(pcDNA3.1(+)-PRSS22)及对照载体(pcDNA3.1(+))转入乳腺癌细胞,实时定量PCR显示pcDNA3.1(+)-PRSS22可明显上调PRSS22在乳腺癌细胞MDA-MB-231及MDA-MB-468中的表达(图6)。
如图7所示,迁移浸润实验结果表明,PRSS22过表达可显著促进乳腺癌细胞MDA-MB-231及MDA-MB-468的迁移浸润能力(图7)。
以上所述仅为本发明的优选实施例而已,并不用于限制本发明,对于本领域的技术人员来说,本发明可以有各种更改和变化。凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (10)
1.一种乳腺癌诊断和/或预后判断用mRNA标志物,其特征在于,所述mRNA标志物为PRSS22。
2.如权利要求1所述标志物,其特征在于,所述标志物选自受试者乳腺癌组织。
3.检测权利要求1或2所述标志物的物质在制备乳腺癌诊断和/或预后产品中的应用。
4.如权利要求3所述的应用,其特征在于,待测样本为人源样本;优选的,所述待测样本为乳腺癌组织。
5.检测PRSS22的试剂组合在制备乳腺癌诊断和/或预后产品中的应用。
6.如权利要求5所述的应用,其特征在于,所述产品包括试剂盒。
7.如权利要求5所述的应用,其特征在于,检测样本为患者的临床生物样本,包括组织、器官中的一种或几种。
8.如权利要求7所述的应用,其特征在于,所述检测样本为乳腺癌组织。
9.如权利要求5所述的应用,其特征在于,所述试剂组合中至少包括如下试剂:
(1)引物;
(2)样本组织裂解试剂;
(3)PCR反应体系。
10.如权利要求9所述的应用,其特征在于,所述引物包括PRSS22引物;所述PRSS22引物序列如下:
PRSS22-F:TCGACAGCCATCCTCAATGC(SEQ ID NO.1);
PRSS22-R:GTGGGTCCCATTCTTCTGGASEQ ID NO.2);
GAPDH-F:GCACCGTCAAGGCTGAGAAC(SEQ ID NO.3);GAPDH-R:TGGTGAAGACGCCAGTGGA(SEQID NO.4)。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2022110117995 | 2022-08-23 | ||
CN202211011799.5A CN115418402A (zh) | 2022-08-23 | 2022-08-23 | 一种乳腺癌诊断和/或预后判断用mRNA标志物及应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN117210563A true CN117210563A (zh) | 2023-12-12 |
Family
ID=84197797
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202211011799.5A Pending CN115418402A (zh) | 2022-08-23 | 2022-08-23 | 一种乳腺癌诊断和/或预后判断用mRNA标志物及应用 |
CN202311062278.7A Pending CN117210563A (zh) | 2022-08-23 | 2023-08-22 | 一种乳腺癌诊断和/或预后判断用mRNA标志物及应用 |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202211011799.5A Pending CN115418402A (zh) | 2022-08-23 | 2022-08-23 | 一种乳腺癌诊断和/或预后判断用mRNA标志物及应用 |
Country Status (1)
Country | Link |
---|---|
CN (2) | CN115418402A (zh) |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007140907A1 (en) * | 2006-06-07 | 2007-12-13 | Bayer Healthcare Ag | Use of serine endopeptidases 22 (prss22) as a therapeutic or diagnostic target |
WO2013106913A1 (en) * | 2012-01-19 | 2013-07-25 | University Health Network | Biomarkers for breast cancer prognosis and treatment |
WO2020056162A1 (en) * | 2018-09-12 | 2020-03-19 | Oregon Health & Science University | Detecting and/or subtyping circulating hybrid cells that correlate with stage and survival |
EP3657170A1 (en) * | 2018-11-20 | 2020-05-27 | Philipps-Universität Marburg | Method for the early detection and diagnosis of primary and relapsed ovarian carcinoma (oc) |
CN111218509B (zh) * | 2019-12-23 | 2023-07-04 | 西南医科大学附属医院 | 乳腺癌的新诊断标志物ppef1及其应用 |
-
2022
- 2022-08-23 CN CN202211011799.5A patent/CN115418402A/zh active Pending
-
2023
- 2023-08-22 CN CN202311062278.7A patent/CN117210563A/zh active Pending
Also Published As
Publication number | Publication date |
---|---|
CN115418402A (zh) | 2022-12-02 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Bachtiary et al. | Gene expression profiling in cervical cancer: an exploration of intratumor heterogeneity | |
Lu et al. | SATB1 is an independent prognostic marker for gastric cancer in a Chinese population | |
Borczuk et al. | Lung adenocarcinoma global profiling identifies type II transforming growth factor-β receptor as a repressor of invasiveness | |
JP2008536488A (ja) | 遺伝子発現署名を使用する、癌による死亡および前立腺癌生存率を予測するための方法および組成物 | |
Chen et al. | Melanoma long non-coding RNA signature predicts prognostic survival and directs clinical risk-specific treatments | |
CN106755344A (zh) | 用于胰腺癌临床预后诊断的分子标记物及其应用 | |
EP3332027A1 (en) | Method for the prediction of progression of bladder cancer | |
WO2018219264A1 (zh) | 一种长链非编码rna作为前列腺癌分子标志物的应用 | |
WO2020157070A1 (en) | Novel biomarkers and diagnostic profiles for prostate cancer | |
JP2010537658A (ja) | Her2+患者におけるガンの予後判定のための方法およびツール | |
CN114410785A (zh) | hsa_circ_0003045作为乳腺癌诊断和/或预后标志物的应用 | |
Halawa et al. | The role of liquid biopsy in the diagnosis and prognosis of WHO grade 4 astrocytoma | |
US20190316205A1 (en) | Mmp1 gene transcript for use as a marker for diagnosis of ovarian cancer prognosis, and test method | |
CN110331207A (zh) | 肺腺癌生物标志物及相关应用 | |
Dadiani et al. | Tumor evolution inferred by patterns of microRNA expression through the course of disease, therapy, and recurrence in breast cancer | |
US20130260384A1 (en) | Method for determining cancer prognosis and prediction with cancer stem cell associated genes | |
CN117210563A (zh) | 一种乳腺癌诊断和/或预后判断用mRNA标志物及应用 | |
Kondoh et al. | Gene expression signatures that classify the mode of invasion of primary oral squamous cell carcinomas | |
Osei et al. | A review of current clinical biomarkers for prostate cancer: towards personalised and targeted therapy | |
CN114908171B (zh) | 人HHIPL2 mRNA在非小细胞肺癌靶向治疗和预后评估中的应用及试剂盒 | |
CN110592218B (zh) | 一种诊治乳腺癌的生物标志物 | |
CN114994336B (zh) | GrB在预测胃癌预后或免疫特征的应用 | |
CN117305452B (zh) | 人SNRK mRNA在非小细胞肺癌诊断、靶向治疗和预后评估中的应用及试剂盒 | |
CN110541034B (zh) | Linc01992在乳腺癌诊疗中的应用 | |
CN109762897B (zh) | 一种骨肉瘤生物标记物环状RNA-circ_0006633及其应用 |
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 |