TWIST1基因通过调节mTOR信号通路对口腔鳞癌Tca8113细胞增殖凋亡的影响  

Effect of TWIST1 gene on proliferation and apoptosis of oral squamous cell carcinoma Tca8113 by regulating mTOR signaling pathway

作　　者：程珏 辛鹏飞 程琳 庄润涛 刘志荣 CHENG Jue;XIN Peng-Fei;CHENG Lin;ZHUANG Run-Tao;LIU Zhi-Rong(Beijing Jiaotong University Community Health Services Center,Beijing 100044,China)

机构地区：[1]北京交通大学社区卫生服务中心,北京100044 [2]山西白求恩医院,太原030032

出　　处：《中国免疫学杂志》2021年第12期1434-1438,共5页Chinese Journal of Immunology

摘　　要：目的:研究TWIST1基因对口腔鳞癌细胞增殖、凋亡的影响以及mTOR信号通路的调控作用。方法:构建shTWIST1质粒(sh-TWIST1)和阴性对照质粒(shRNA),利用Lipofectamine 3000转染试剂分别将构建好的两种质粒转染至口腔鳞癌Tca8113细胞中,以只加入转染试剂的Tca8113细胞作为空白对照(Control)。QRT-PCR法检测Tca8113细胞中TWIST1mRNA水平;CCK-8法检测TWIST1对Tca8113细胞活力的影响;Edu染色法检测TWIST1对Tca8113细胞增殖的影响;TUNEL染色法检测TWIST1对Tca8113细胞凋亡的影响;ELISA法检测TWIST1对Tca8113细胞中Beclin1、LC3水平的影响;Western blot法检测TWIST1对Tca8113细胞中p-mTOR和P-P70S6K蛋白表达情况的影响。结果:与阴性对照组和空白对照组相比,shTWIST1质粒组Tca8113细胞中TWIST1 mRNA水平下降,细胞活力和增殖能力明显提高,其凋亡能力随之下降;Tca8113细胞中Beclin1水平下降、LC3水平升高;且p-mTOR和P-P70S6K蛋白升高;而阴性对照组与空白对照组之间无差异。结论:TWIST1沉默后过度激活Tca8113细胞自噬反应,导致Tca8113细胞增殖,并抑制凋亡,其机制可能与激活mTOR信号通路有关。Objective:To study the effect of TWIST1 gene on the proliferation and apoptosis of oral squamous carcinoma cells,and the regulation of mTOR signaling pathway.Methods:The sh-TWIST1 plasmid(sh-TWIST1)and negative control plasmid(shRNA)was constructed,two plasmids were transfected into oral squamous cell carcinoma Tca8113 cells by using Lipofectamine 3000 transfection reagent,and Tca8113 cells with Lipofectamine 3000 transfection reagent as control group. The QRT-PCR method was used to detect TWIST1 mRNA levels in Tca8113 cells;the CCK-8 method was used to detect the effect of TWIST1 on the viability of Tca8113 cells;the Edu staining method was used to detect the effect of TWIST1 on the proliferation of Tca8113 cells;the TUNEL staining was used to detect the effect of TWIST1 on the apoptosis of Tca8113 cells;ELISA method was used to detect the levels of Beclin1 and LC3 on Tca8113 cells;Western blot method was used to detect the expression of p-mTOR and P-P70 S6 K protein on Tca8113 cells.Results:Compared with the negative control group and the blank control group,the TWIST1 mRNA level in the Tca8113 cells of the sh-TWIST1 plasmid group was decreased;the cell viability and proliferative ability were significantly improved;and the apoptosis ability decreased;the Beclin1 level in Tca8113 cells was down-regulated and the LC3 level was increased;and the expression of p-mTOR and P-P70 S6 K protein in Tca8113 cells was increased;but there was no difference between the negative control group and the blank control group.Conclusion:After TWIST1 silencing,excessive activation of autophagy in Tca8113 cells leads to proliferation of Tca8113 cells and the inhibition of cells apoptosis,the underlying mechanism is related to activation of mTOR signaling pathway.

关 键 词：TWIST1 口腔鳞癌 自噬 增殖凋亡 MTOR信号通路 

分 类 号：R739.85[医药卫生—肿瘤]