应用基因芯片研究E2F-1过表达对胃癌细胞免疫相关基因表达的影响

Effects of E2F-1 overexpression on immunity-relative genes of gastric carcinoma cells by gene chip

目的利用基因表达谱芯片技术筛选过表达E2F-1的胃癌MGC-803细胞与对照组间差异表达的免疫相关基因,初步探讨E2F-1对胃癌细胞免疫功能影响的机制。方法分别抽取转染E2F-1的胃癌MGC-803细胞和未转染E2F-1的胃癌MGC-803细胞的总RNA.纯化mRNA,逆转录合成cDNA标记后,与22K人类基因表达谱芯片进行杂交,扫描后筛选出与免疫相关的差异表达基因,半定量RT-PCR方法验证差异表达基因。结果在21522条基因中,两组细胞间差异表达的免疫相关基因有10条,其中上调基因2条,下调基因8条。上调基因中有1条功能信息不明。其中myc的RT-PCR结果与基因芯片扫描结果相似,具有相同的方向性。结论利用基因芯片技术筛选出E2F-1过表达影响胃癌细胞免疫功能的相关基因,为今后更深入的研究E2F-1对胃癌免疫功能的影响奠定了基础。

Objective To investigate the effects of E2F-1 overexpression on immunity-relative genes of gastric carcinoma MGCS03 cells by cDNA microarray technique and to explore the mechanism of E2F-1 affecting immune functions of gastric cancer. Methods The total RNA was extracted from gastric cancer cell line MC, C-803 transfected with E2F-1 or those control cells without transfection and then purified. The cDNA was obtained by reverse tran- scription polymerase chain reaction （RT-PCR）, and then labeled with fluorescence as probes, which were hybridized with gene chip containing human 22K gene expression profile. Subsequently, the signal images were scanned by LuxScan 10K/A dual pathways laser scanner and analyzed by LuxScan3.0 image analysis software. The semi-quantitative RT-PCR was used to confirm the array results. Results Of the 21522 target genes, 10 genes were screened out for differences in immunity-relative gene expression level. Two of the 10 genes were up-regulated and 7 were down-regulated. The function of a differentially expressed gene was unknown. The result of RT-PCR was coincided well with the cDNA microarray results. Conclusion Ten immunity-relative genes in gastric cancer cells are found by using gene chip, which provides a basis for further investigation of the effects of E2F-1 on immune function of gastric cancer.