基于蛋白质组学的乳腺癌标志物筛选及其应用

Screening breast cancer biomarkers through proteomics analysis

目的利用高通量质谱技术筛选不同亚型的乳腺癌发生发展及预后标志物,有助于建立更有效的乳腺癌诊断手段、预测病因及干预治疗。方法选取4例乳腺癌患者标本,包括4例癌组织和4例对应癌旁组织,采用SDT裂解法提取蛋白质,并BCA法进行蛋白质定量后,进行肽段的脱盐及定量。随后进行色谱分级、液相色谱-串联质谱(liquid chromatography-tandem mass spectrometry,LS-MS/MS)数据采集。用软件MaxQuant进行查库鉴定及定量分析。提取样本RNA经qPCR筛选出乳腺癌差异表达标志物。结果分析4例乳腺癌样本的蛋白质组定量分析数据最后筛选出LCP1、ARF1、CCT3、VDAC1、NANS、NME1等23个癌组织中特异性表达的蛋白。且将相关蛋白所对应的基因与其癌旁进行比较筛选出在侵袭性乳腺癌癌组织中表达升高的23个基因。qPCR的结果表明最后有14个基因差异有统计学意义。结论通过筛选及分析包括以下一种:LCP1、CCT3、NANS、NME1、HSP90AA1、RAN、YWHAZ、ERH、PRDX1、STIP1、HSPA8、H2AFY、HSPE1和PDIA3标志物。利用这些标志物在乳腺癌癌组织及癌旁组织的差异表达,可用于乳腺癌诊断、预后评估。

Objective To use high-throughput mass spectrometry screening of different subtypes of breast cancer markers about it’s development and prognosis.For patients with different subtypes,Which established more effective breast cancer diagnosis methods,predict the pathogen and intervention treatment.Methods Selected 4 samples of breast cancer patients,included 4 cancer and paracancerous tissues,the protein was extracted by SDT lysis method and the protein was quantified by BCA method then desalt and quantify peptides.Thechromatographic classification and liquid chromatography-tandem mass spectrometry(LS-MS/MS)to collect datawasutilized.The software Max-Quant for library identification and quantitative analysis was uesd.Finally,the sample RNA and the differentially expressed markers of breast cancer were screened after qPCR was seleted.Results Quantitative data from the proteome of 4 breast cancer samples showed that compared with paracancerous,23 specific proteins expressed in cancer tissues were seleced.The proteins corresponding to genes associated therewith paracancerous comparing selected tissue had elevated expression in invasive breast cancer 23 genes with significant difference.The results of qPCR showed that 14 genes were significantly different in the end.Conclusion By selection and comprises at least one:LCP1,CCT3,NANS,NME1,HSP90AA1,RAN,YWHAZ,ERH,PRDX1,STIP1,HSPA8,H2AFY,HSPE1 and PDIA3 markers.The differential expression of these markers in breast cancer tissues and adjacent tissues can be used for breast cancer diagnosis,prognostic evaluation or drug screening.