论文部分内容阅读
目的:研究带RGD的腺病毒介导的ING4对人鼻咽癌细胞CNE生长、凋亡及细胞周期的影响并探讨其可能调节机制。方法:以Ad.RGD-ING4及腺病毒空载体感染CNE细胞,RT-PCR法检测ING4基因的表达水平,Western blot法检测目的蛋白的表达。用MTT试验检测ING4对CNE细胞生长情况的影响,用Annexin V-PE/7-AAD双染色测定ING4对CNE细胞凋亡的影响,用PI单染色检测ING4对CNE细胞细胞周期的影响。RT-PCR检测p21、Bcl-2、Bax基因的表达水平的差异,Western blot法检测Survivin蛋白、Cleaved-caspase3蛋白表达的差异。结果:CNE细胞感染Ad.RGD-ING4 72h后,ING4在CNE细胞中过表达,CNE细胞的生长受到明显抑制,细胞凋亡率明显升高,G2/M期出现明显阻滞。RT-PCR结果显示,感染Ad.RGD-ING4的CNE细胞中Bcl-2表达下降,p21、Bax表达升高,差异具有统计学意义(均P<0.01)。Western blot结果显示Survivin蛋白表达下降,Cleaved-caspase3蛋白表达升高。结论:Ad.RGD-ING4可以对鼻咽癌细胞株CNE的起到抑癌增效作用,这一作用可能是通过下调Bcl-2、Survivin表达及上调p21、Bax、caspase3实现的。
OBJECTIVE: To study the effect of RGD-mediated adenovirus-mediated ING4 on CNE cell growth, apoptosis and cell cycle in human nasopharyngeal carcinoma cells and to explore its possible regulatory mechanism. Methods: CNE cells were infected with Ad.RGD-ING4 and adenovirus empty vector. The expression of ING4 gene was detected by RT-PCR and the expression of target protein was detected by Western blot. The effect of ING4 on the growth of CNE cells was detected by MTT assay. The effect of ING4 on the apoptosis of CNE cells was detected by Annexin V-PE / 7-AAD double staining. The effect of ING4 on the cell cycle of CNE cells was detected by PI single staining. The difference of p21, Bcl-2 and Bax gene expression levels was detected by RT-PCR. The protein expression of Survivin and Cleaved-caspase3 were detected by Western blot. Results: After infected with CNE cells for 72 hours, ING4 was overexpressed in CNE cells. The growth of CNE cells was significantly inhibited. The apoptosis rate was significantly increased and G2 / M phase was significantly blocked. The results of RT-PCR showed that the expression of Bcl-2 and p21, Bax in CNE cells infected with Ad.RGD-ING4 were significantly increased (all P <0.01). Western blot results showed that Survivin protein expression decreased Cleaved-caspase3 protein expression. CONCLUSION: Ad.RGD-ING4 can inhibit the proliferation of nasopharyngeal carcinoma cell line CNE. This effect may be through the down-regulation of Bcl-2, Survivin expression and up-regulation of p21, Bax and caspase3.