目的:利用全基因组表达谱芯片对人肺腺癌和癌旁组织中差异表达的mRNA进行检测,对mRNA在肺腺癌发生过程中可能涉及到的信号通路变化进行分析。方法:提取6例人肺腺癌与癌旁组织标本中的总RNA,体外逆转录制备ds-cDNA并用Cy3荧光标记,与含有19 000条mRNA的芯片进行杂交,扫描软件扫描芯片荧光信号图像,筛选差异表达的mRNA并进行信号通路分析。结果:1差异表达的mRNA总共有832条,其中在肿瘤组织中相对高表达的mRNA有407条,相对低表达的mRNA有425条(大于2倍变化且P〈0.05)。2信号通路分析发现差异表达的mRNA参与了9个信号通路的调控,其中同源重组信号通路、人转化生长因子-β通路、丝裂原激活蛋白激酶信号通路、细胞周期信号通路、p53信号通路等5条信号通路与肿瘤的发生有关。结论:人肺腺癌组织中有大量mRNA发生差异表达,差异表达的mRNA在多个与肺腺癌发生密切相关的信号通路中发挥重要的调控作用。
Objective. To detect the difference of mRNA expression profiling between human pulmonary adenocarcinoma and paraneoplastic tissues and identify the changes in signaling pathways by whole genome expression microarray screening. Methods: Six samples of total RNA from human pulmonary adenocarcinoma and paraneoplastic tissues were extracted, mRNA was reversely tran- scripted into ds-cDNA in vitro and fluorescently labeled with Cy3 then hybridized with chips con- taining 19 000 mRNAs. The chip fluorescence signal image was scanned to determine the differ- ential expression of mRNAs and the changes in signaling pathways. Results: ①Among the 19 000 mRNAs, 832 were differentially expressed in pulmonary adenocarcinoma tissues, in which 407 mRNAs were up-regulated and 425 mRNAs were down-regulated compared with the paraneoplas- tie tissues (2 fold changes and P〈0.05). ②Nine signaling pathways were involved based on mR- NAs expression data and 5 of which were involved in tumorigenesis (homologous recombination,TGF-β signaling pathway, MAPK signaling pathway, cell cycle and p53 signaling pathway). Conclusion: ①Obvious changes of mRNAs expression profiling were observed in the human pul- monary adenocarcinoma. ②Differentially expressed mRNAs play an important role in regulating signaling pathways in the human pulmonary adenocarcinoma tumorgenesis.