Ad.RGD-ING4对人鼻咽癌细胞CNE抑癌增效及其机制的研究

发布时间：2018-03-18 20:41

本文选题：Ad.RGD-ING　切入点：鼻咽肿瘤　出处：《临床耳鼻咽喉头颈外科杂志》2014年06期 　论文类型：期刊论文

【摘要】：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P0.01)。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。
[Abstract]:Objective: to study the effect of adenovirus mediated ING4 with RGD on CNE growth of human nasopharyngeal carcinoma cells. Methods: Ad.RGD-ING4 and adenovirus empty vector infected CNE cells were used to detect the expression level of ING4 gene by RT-PCR and Western blot to detect the expression of target protein. ING4 was detected by MTT assay. On the growth of CNE cells, The effect of ING4 on apoptosis of CNE cells was determined 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 the expression level of p21, Bcl-2 and Bax gene was detected by RT-PCR. The expression of Cleaved-caspase-3 protein in CNE cells was detected by Western blot. Results after Ad.RGD-ING4 72 hours after Ad.RGD-ING4 infection, the expression of ING4 was overexpressed in CNE cells. Cell growth was significantly inhibited, The results of RT-PCR showed that the expression of Bcl-2 in CNE cells infected with Ad.RGD-ING4 decreased and the expression of p21 + Bax increased. The difference was statistically significant (P0.01N. Western blot showed that the expression of Survivin protein decreased and Cleaved-caspase3 protein increased. Conclusion: Ad.RGD-ING4 can inhibit the cancer of nasopharyngeal carcinoma cell line CNE, and this effect may be achieved by down-regulating the expression of Bcl-2G survivin and upregulating p21 Baxcaspase3.
【作者单位】：苏州大学附属第一医院耳鼻咽喉科;苏州大学基础医学院细胞与分子生物学教研室;
【基金】：江苏省苏州市国标技术引进与合作创新项目(No:SH201209)
【分类号】：R739.63

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