PCNA对口腔鳞癌细胞凋亡和增殖的影响

Effect of PCNA on apoptosis and proliferation of oral squamous cell carcinoma TCA8113 cells

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。

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 cytometry, and the effects of PCNA hyper expression or gene silence on cell apoptosis were analyzed. Results PCNA hyper expression and down-regulation plasmid were constructed and transfected into TCA8113 cells successfully. The PCNA hyper expression and down-regulation were verified by Western blotting and Realtime PCR, respectively. MTT proliferation assay revealed that the PCNA hyper expression promoted TCA8113 cell proliferation. The survival rates of PCNA over-expressed cells at 24, 48 and 72 h were significantly higher than those of control cells（P〈0.05）. PCNA silence effectively inhibited TCA8113 cell proliferation. The survival rates of PCNA down-regulated cells at 24, 48 and 72 h were significantly lower than those of control cells（P〈0.05）. By treating with cisplatin, it was found that PCNA hyper expression inhibited cell apoptosis obviously, while PCNA silence facilitated cell apoptosis significantly. Conclusion PCNA gene is involved in the apoptosis and proliferation of OSCC TCA8113 cells, and it can significantly enhance the sensitivity of TCA8113 to chemotherapeutic drug cisplatin.