三阴性与三阳性乳腺癌定量蛋白质组学和生物信息学比较研究
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摘要
目的雌/孕激素受体(estrogen receptor/progesterone receptor,ER/PR)和癌基因人表皮生长因子受体2(human epidermal growth factorreceptor2,HER2)与乳腺癌的发生和发展密切相关,其表达状态是乳腺癌分子分型、治疗方案选择和预后预测的重要依据。为了深入探讨ER/PR/HER2在乳腺癌发生和恶性演进中的意义及相关机制,拟建立ER/PR/HER-2阴性和阳性乳腺癌的蛋白质表达谱,寻找ER/PR/HER-2阴性和阳性乳腺癌中差异表达蛋白,为乳腺癌患者提供新的预后预测指标和治疗新靶点。方法应用蛋白质组学同位素相对标记与绝对定量(isobaric tags for relative and absolute quantitation,iTRAQ)技术建立ER/PR/HER-2阳性和阴性乳腺癌的蛋白质差异表达谱,鉴定2组乳腺癌的差异表达蛋白,对部分差异表达蛋白进行生物信息学分析,包括蛋白功能注释、基因本体论分类分析(gene ontology classification analysis,GO分析)和京都基因与基因组百科全书(Kyoto encyclopedia of gene and genome,KEGG)通路分析。结果应用iTRAQ蛋白质组学技术对乳腺癌组织进行了蛋白组学分析,鉴定出ER/PR/HER-2阳性和阴性组间有差异表达的蛋白4 999种,以ER/PR/HER-2阳性∶ER/PR/HER-2阴性≥3为上调标准,确定ER/PR阳性组上调蛋白73种。以ER/PR/HER-2阳性∶ER/PR/HER-2阴性≤0.5为下调标准,ER/PR/HER-2阳性组下调蛋白58种。GO分析结果显示,ER/PR/HER-2阴性和阳性乳腺癌差异表达蛋白的分子功能、生物过程和细胞定位较为复杂,并且在上调蛋白和下调蛋白上存在分布差异。KEGG通路分析发现,部分差异表达蛋白涉及200条信号通路。结论 ER/PR/HER-2阳性和阴性乳腺癌间存在差异表达蛋白,这些蛋白涉及复杂的分子功能、生物过程和信号通路,可能与ER/PR/HER-2的表达状态有关。
OBJECTIVE Estrogen/progesterone receptor(PR)and oncogene HER-2are closely related to breast cancer occurrence and development,its expression is an important basis for the prediction of breast cancer molecular classification,treatment and prognosis.In order to further investigate the significance and related mechanism of the ER/PR/HER-2in malignant progression of breast cancer,this study planed to establish differentially expressed proteins profiles for ER/PR/HER-2negative and positive breast carcinoma,and provide new prognostic markers and therapeutic targets for patients with ER/PR/HER-2positive breast cancer.METHODS By proteomic iTRAQ technology,ER/PR/HER-2positive and negative breast cancer protein expression profiles were established,differences of protein expression were identified and parts of differential expression protein were analyzed by bioinformatics analysis,including protein function annotation and GO classification analysis and KEGG pathway analysis.RESULTS iTRAQ proteomic analysis of breast cancer tissue with identified ER/PR/HER-2positive and negative groups showed 4 999 differentially expressed protein.While ratio of ER/PR/HER-2(+)/ER/PR/HER-2(-)was more than or equal to 3for standard cut,73up-regulated proteins in ER/PR/HER-2positive group were identified.In ER/PR/HER-2(+)/ER/PR/HER-2(-)less than or equal 0.5for standard cut,58down-regulated proteins were identified in ER/PR/HER-2positive group.The results of GO analysis showed that the cellular composition,molecular function,biological process of differentially expressed proteins were complex between ER/PR/HER-2positive and negative breast cancer,and there were differences in the distribution of up-regulated proteins and down-regulation of proteins.KEGG pathway analysis showed that differentially expressed proteins involved in 200 signal pathways.CONCLUSION There are differentially expressed proteins between ER/PR/HER-2positive and negative breast cancer,which involves complex molecular function,biological process and signaling pathway,which may be related to the expression state of ER/PR/HER-2.
OBJECTIVE Estrogen/progesterone receptor(PR)and oncogene HER-2are closely related to breast cancer occurrence and development,its expression is an important basis for the prediction of breast cancer molecular classification,treatment and prognosis.In order to further investigate the significance and related mechanism of the ER/PR/HER-2in malignant progression of breast cancer,this study planed to establish differentially expressed proteins profiles for ER/PR/HER-2negative and positive breast carcinoma,and provide new prognostic markers and therapeutic targets for patients with ER/PR/HER-2positive breast cancer.METHODS By proteomic iTRAQ technology,ER/PR/HER-2positive and negative breast cancer protein expression profiles were established,differences of protein expression were identified and parts of differential expression protein were analyzed by bioinformatics analysis,including protein function annotation and GO classification analysis and KEGG pathway analysis.RESULTS iTRAQ proteomic analysis of breast cancer tissue with identified ER/PR/HER-2positive and negative groups showed 4 999 differentially expressed protein.While ratio of ER/PR/HER-2(+)/ER/PR/HER-2(-)was more than or equal to 3for standard cut,73up-regulated proteins in ER/PR/HER-2positive group were identified.In ER/PR/HER-2(+)/ER/PR/HER-2(-)less than or equal 0.5for standard cut,58down-regulated proteins were identified in ER/PR/HER-2positive group.The results of GO analysis showed that the cellular composition,molecular function,biological process of differentially expressed proteins were complex between ER/PR/HER-2positive and negative breast cancer,and there were differences in the distribution of up-regulated proteins and down-regulation of proteins.KEGG pathway analysis showed that differentially expressed proteins involved in 200 signal pathways.CONCLUSION There are differentially expressed proteins between ER/PR/HER-2positive and negative breast cancer,which involves complex molecular function,biological process and signaling pathway,which may be related to the expression state of ER/PR/HER-2.
引文
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[3]Puig-Vives M,Sánchez MJ,Sánchez-Cantalejo J,et al.Distribution and prognosis of molecular breast cancer subtypes defined by immunohistochemical biomarkers in a Spanich populationbased study[J].Gynecol Oncol,2013,130(3):609-614.
[4]Najaf B,Anvari S,Roshan ZA.Disease free survival among molecular subtypes of early stage breast cancer between 2001and2010in Iran[J].Asian Pac J Cancer Prev,2013,14(10):5811-5816.
[5]Lee O,Choi MR,Christov K,et al.Progesterone receptor antagonism inhibits progestogen-related carcinogenesis and suppresses tumor cell proliferation[J].Cancer Lett,2016,376(2):310-317.
[6]Sun W,Gu C,Xia M,et al.Significance of estrogen receptor subtypes in breast tumorigenesis and progression[J].Tumour Biol,2014,35(9):9111-9117.
[7]Zhang M,Chen H,Gu J.Analysis of factors affecting endocrine therapy resistance in breast cancer[J].Oncol Lett,2016,11(1):379-384.
[8]He ZY,Wu SG,Yang Q,et al.Breast cancer subtype is associated with axillary lymph node metastasis:A retrospective cohort study[J/CD].Medicine(Baltimore),2015,94(48):e2213.
[9]Tsai CH,Chiu JH,Yang CW,et al.Molecular characteristics of recurrent triple-negative breast cancer[J].Mol Med Rep,2015,12(5):7326-7334.
[10]Lambertini M,Santoro L,Del Mastro L,et al.Reproductive behaviors and risk of developing breast cancer according to tumor subtype:A systematic review and meta-analysis of epidemiological studies[J].Cancer Treat Rev,2016,49:65-76.
[11]Clarke R,Tyson JJ,Dixon JM.Endocrine resistance in breast cancerAn overview and update[J].Mol Cell Endocrinol,2015,418(3):220-234.
[12]Kassem L,Deygas M,Fattet L,et al.TIF1γinterferes with TGFβ1/SMAD4signaling to promote poor outcome in operable breast cancer patients[J].BMC Cancer,2015,15:453.
[13]Colavito SA,Zou MR,Yan Q,et al.Significance of glioma-associated oncogene homolog 1(GLI1)expression in claudin-low breast cancer and crosstalk with the nuclear factor kappa-lightchain-enhancer of activated B cells(NFκB)pathway[J].Breast Cancer Res,2014,16(5):444.
[14]Singel SM,Batten K,Cornelius C,et al.Receptor-interacting protein kinase 2promotes triple-negative breast cancer cell migration and invasion via activation of nuclear factor-kappaB and c-Jun N-terminal kinase pathways[J].Breast Cancer Res,2014,16(2):R28.
[15]Kerdivel G,Boudot A,Habauzit D,et al.Activation of the MKL1/actin signaling pathway induces hormonal escape in estrogen-responsive breast cancer cell lines[J].Mol Cell Endocrinol,2014,390(1-2):34-44.
[16]Martin JL,de Silva HC,Lin MZ,et al.Inhibition of insulin-like growth factor-binding protein-3signaling through sphingosine kinase-1sensitizes triple-negative breast cancer cells to EGF receptor blockade[J].Mol Cancer Ther,2014,13(2):316-328.
[17]Chen WC,Lai YA,Lin YC,et al.Curcumin suppresses doxorubicin-induced epithelial-mesenchymal transition via the inhibition of TGF-βand PI3K/AKT signaling pathways in triple-negative breast cancer cells[J].J Agric Food Chem,2013,61(48):11817-11824.
[18]Bilir B,Kucuk O,Moreno CS.Wnt signaling blockage inhibits cell proliferation and migration,and induces apoptosis in triplenegative breast cancer cells[J].J Transl Med,2013,11:280.
[19]Mukherjee N,Bhattacharya N,Alam N,et al.Subtype-specific alterations of the Wnt signaling pathway in breast cancer:clinical and prognostic significance[J].Cancer Sci,2012,103(2):210-220.
[20]Fearon AE,Gould CR,Grose RP.FGFR signalling in women’s cancers[J].Int J Biochem Cell Biol,2013,45(12):2832-2842.
[21]Zhang X,Mu X,Huang O,et al.Luminal breast cancer cell lines overexpressing ZNF703are resistant to tamoxifen through activation of Akt/mTOR signaling[J/CD].PLoS One,2013,8(8):e72053.
[22]Lian WJ,Liu G,Liu YJ,et al.Downregulation of BMP6enhances cell proliferation and chemoresistance via activation of the ERK signaling pathway in breast cancer[J].Oncol Rep,2013,30(1):193-200.
[23]Liu RZ,Graham K,Glubrecht DD,et al.A fatty acid-binding protein7/RXRβpathway enhances survival and proliferation in triple-negative breast cancer[J].J Pathol,2012,228(3):310-321.
[24]Boulaiz H,Alvarez PJ,Prados J,et al.gef gene expression in MCF-7breast cancer cells is associated with a better prognosis and induction of apoptosis by p53-mediated signaling pathway[J].Int J Mol Sci,2011,12(11):7445-7458.
[25]Li H,Yang B,Huang J,et al.Naringin inhibits growth potential of human triple-negative breast cancer cells by targetingβ-catenin signaling pathway[J].Toxicol Lett,2013,220(3):219-228.