PCNA对口腔鳞癌细胞凋亡和增殖的影响.PDF

Med J Chin PLA, Vol. 39, No. 10, October 1, 2014 　795 论　著 PCNA对口腔鳞癌细胞凋亡和增殖的影响 赵莉琳，刘阳 [摘要]　目的　探讨增殖细胞核抗原(PCNA)基因对口腔鳞癌细胞TCA8113增殖和凋亡的影响。方法　 构建PCNA 高表达以及RNA干扰载体，分别采用Western blotting 或定量PCR检测其表达水平。将其转染TCA8113 口腔鳞癌细胞使 PCNA高表达或沉默后，采用MTT实验检测PCNA对TCA8113细胞增殖的影响，通过AnnexinV/PI染色、流式细胞仪检 测细胞凋亡情况，分析PCNA高表达或沉默对细胞凋亡的影响。结果　成功构建PCNA高表达和PCNA干扰载体，分 别转染TCA8113细胞后，Western blotting 和定量PCR检测到PCNA 的高表达或低表达。MTT增殖实验结果显示，PCNA 高表达可促进TCA8113细胞增殖，24 、48 、72h 时PCNA高表达组细胞的存活率均明显高于对照组(P 0.05) 。PCNA沉 默可有效抑制TCA8113细胞增殖，24 、48 、72h时PCNA沉默组细胞的存活率均明显低于对照组(P 0.05) 。在顺铂诱导 细胞凋亡过程中，PCNA高表达可明显抑制凋亡，PCNA沉默则显著促进凋亡。结论　PCNA基因参与口腔鳞癌细胞 TCA8113 的增殖和凋亡，能显著增强TCA8113对化疗药物顺铂的敏感性。 [关键词]　口腔肿瘤；TCA8113细胞；增殖细胞核抗原；顺铂；RNA干扰 [中图分类号]　R78　　　　[文献标志码]　A　　　　[文章编号]　0577-7402(2014)10-0795-05 [DOI]　10.11855/j.issn.0577-7402.2014.10.07 Effect of PCNA on apoptosis and proliferation of oral squamous cell carcinoma TCA8113 cells ZHAO Li-lin1, LIU Yang2* 1Department of Stomatology, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China 2 Otolaryngology Head and Neck Surgery, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China * Corresponding author, E-mail: liuyanghhhh@163.com [Abstract]　Objective　To explore the effect of proliferating cell nuclear antigen (PCNA) on apoptosis and proliferation of oral squamous cell carcinoma (OSCC) TCA8113 cells. Methods 　Hyper expressed plasmid pcDNA3.1-PCNA and RNA interference (RNAi) vector pSilencer-shPCNA were constructed and transfected into TCA8113 cells, and their expression was determined by Western blotting or Real-time PCR. The cell proliferation was then assessed using MTT assay. Cell apoptosis was detected by Annexin V/PI staining and flow cytom etry, and the effects of PCNA hyper expression or gene silence on cell apoptosis were analyzed. Results　PCNA hyper expression and down-regulation pl