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目的探讨卵巢癌细胞系SKOV3稳定表达程序性细胞死亡4(PDCD4)基因后基因表达谱的变化。方法将pDsRed2-N1-PDCD4真核表达载体和pDsRed2-N1空载体分别转染至SKOV3并获得稳定细胞系,应用基因芯片技术分析PDCD4基因转染后基因表达的变化,RT-PCR检测基因的表达以验证芯片结果。结果PDCD4转染SKOV3后引起大量基因表达的变化,表达明显差异基因共有467个,其中上调255个,下调212个;RT-PCR验证选取的5个基因的表达差异和芯片显示的结果一致。结论成功筛选出PDCD4基因转染卵巢癌细胞系SKOV3后的差异表达基因,为进一步研究PDCD4的抑癌作用机制提供了实验依据。
Objective To investigate the changes of gene expression profile after stably expressing PDCD4 gene in ovarian cancer cell line SKOV3. Methods The pDsRed2-N1-PDCD4 eukaryotic expression vector and pDsRed2-N1 empty vector were transfected into SKOV3 cells respectively to obtain stable cell lines. The gene expression changes of PDCD4 gene were analyzed by gene chip technology. The expression of PDCD4 gene was detected by RT-PCR To verify chip results. RESULTS: A total of 467 differentially expressed genes were found in SKOV3 cells transfected with PDCD4, of which 255 were up-regulated and 212 were down-regulated. Differences in the expression of 5 genes tested by RT-PCR were in accordance with those indicated by the chips. Conclusion The differentially expressed genes of PDCD4 gene transfected ovarian cancer cell line SKOV3 were successfully screened, which provided an experimental basis for further study on the mechanism of PDCD4 tumor suppressor.