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旨在研究携带人IL-24基因的腺病毒表达载体(Ad-IL-24)对SGC-7901人胃癌细胞的生长抑制作用并分析其分子机制。以不同MOI(感染复数)的Ad空载体腺病毒感染SGC-7901人胃癌细胞,筛选出最佳感染剂量;Ad-IL-24以最佳感染剂量感染SGC-7901胃癌细胞,RT-PCR法和Western blotting法检测腺病毒介导的IL-24基因在SGC-7901胃癌细胞中的转录;MTT法检测Ad-IL-24对SGC-7901胃癌细胞的生长抑制作用,流式细胞仪检测其诱导SGC-7901人胃癌细胞凋亡和细胞周期改变的效应,Hoechst33258染色荧光显微镜检测其诱导胃癌细胞凋亡的核形态改变;RT-PCR半定量法进一步检测SGC-7901胃癌细胞中凋亡相关基因的转录。结果显示,100MOI为感染SGC-7901胃癌细胞的腺病毒最佳感染剂量;Ad-IL-24能成功介导IL-24基因在SGC-7901胃癌细胞中转录性表达;Ad-IL-24感染SGC-7901胃癌细胞后,能明显抑制胃癌细胞生长和诱导凋亡;Ad-IL-24能显著上调SGC-7901胃癌细胞中bax、caspase-3和p53的表达和下调bcl-2的表达。因此,腺病毒介导的IL-24表达具有明显抑制SGC-7901人胃癌细胞生长和诱导凋亡的抗肿瘤效应,其机制可能与上调bax/bcl-2、caspase-3和p53密切相关。
The purpose of this study was to investigate the growth inhibitory effect of adenovirus vector carrying human IL-24 gene (Ad-IL-24) on SGC-7901 human gastric cancer cells and analyze its molecular mechanism. Ad-IL-24 was transfected into SGC-7901 human gastric cancer cells with Ad-IL-24 at different MOI (multiplicity of infection) to infect SGC-7901 human gastric cancer cells. The optimal infection dose was obtained by Ad-IL- The transcription of adenovirus-mediated IL-24 gene in SGC-7901 gastric cancer cells was detected by Western blotting. The growth inhibition of SGC-7901 gastric cancer cells by Ad-IL-24 was detected by MTT assay. -7901 human gastric cancer cell apoptosis and cell cycle changes in the role of Hoechst33258 staining fluorescence microscopy to detect apoptosis of gastric cancer cell nuclear morphological changes; semi-quantitative RT-PCR method to further detect SGC-7901 gastric cancer cell apoptosis-related gene transcription . The results showed that 100 MOI was the optimal infection dose of adenovirus for SGC-7901 gastric cancer cells; Ad-IL-24 could successfully mediate the transcriptional expression of IL-24 gene in SGC-7901 gastric cancer cells; Ad-IL- -7901 gastric cancer cells can significantly inhibit the growth of gastric cancer cells and induce apoptosis; Ad-IL-24 can significantly up-regulate the expression of bax, caspase-3 and p53 and down-regulate the expression of bcl-2 in SGC-7901 gastric cancer cells. Therefore, adenovirus-mediated IL-24 expression can significantly inhibit the growth of SGC-7901 human gastric cancer cells and induce apoptosis of the anti-tumor effect, the mechanism may be upregulated bax / bcl-2, caspase-3 and p53 are closely related.