干扰TAZ基因对舌鳞癌细胞CAL27增殖凋亡的影响及其机制

doi:10.6040/j.issn.1671-7554.0.2017.985

摘要/Abstract

摘要： 目的探讨干扰TAZ基因后对舌鳞癌细胞CAL27增殖和凋亡的影响以及可能的分子调控机制。方法实验分为干扰组和对照组,将稳定干扰TAZ基因的慢病毒载体转染至舌鳞癌细胞CAL27中作为干扰组,并以非特异性序列转染CAL27细胞作为对照组。利用实时荧光定量PCR(RT-PCR)及Western blotting检测干扰效率,CCK-8细胞活力实验及EdU染色法检测转染后细胞增殖能力的改变,流式细胞术检测细胞周期和细胞凋亡情况,Western blotting检测与周期及凋亡相关蛋白的表达情况。结果干扰TAZ基因后,RT-PCR和Western blotting检测结果显示,TAZ mRNA和蛋白表达水平明显降低;CCK-8细胞增殖活力检测及EdU染色法显示细胞增殖能力降低;细胞早期凋亡和晚期凋亡比例均增多; G0/G1期细胞比例增多,而G2/M期细胞减少。同时,干扰TAZ基因后,周期相关蛋白CYCLIN D1、CDK4和CDK6等表达水平降低;而凋亡相关蛋白Bax、cleaved Caspase9表达水平均升高,Bcl-2表达水平降低。结论干扰TAZ基因可以通过调控细胞周期和凋亡相关蛋白的表达,来影响舌鳞癌细胞的增殖和凋亡。

关键词: TAZ基因, 舌鳞癌细胞CAL27, 细胞增殖, 细胞凋亡

Abstract: Objective To explore the effects of TAZ knockdown on the proliferation and apoptosis of tongue squamous cell carcinoma(TSCC)cell line CAL27, and the possible molecular mechanism. Methods CAL27 cells were divided into the interference group and control group. The former was transfected with targeting TAZ shRNA, and the latter with non-specific targeting sequence. The interference efficiency was detected with real-time fluorescent quantitative PCR(RT-PCR)and Western blotting. The changes of proliferation were detected with CCK-8 assay and EdU staining assay. The cell cycle distribution and cell apoptosis were tested with flowcytometry. The expressions of proteins related to cell cycle and apoptosis were determined with Western blotting. Results The results of RT-PCR and Western blotting showed 山东大学学报 (医学版)56卷10期 -陈希彦,等.干扰TAZ基因对舌鳞癌细胞CAL27增殖凋亡的影响及其机制 \=-that the mRNA and protein expressions of the interference group decreased sufficiently. The CCK-8 and EdU staining assay indicated that knockdown of TAZ led to the inhibition of proliferation activity. Flowcytometry results showed that cell cycle arrested in G0/G1 phase and cell apoptosis increased after TAZ knockdown. Meanwhile, knockdown of TAZ decreased the expressions of CYCLIN D1, CDK4, and CDK6. By contrast, the expressions of proapoptotic proteins like Bax and cleaved Caspase9 increased, while the expression of anti-apoptotic protein like Bcl-2 decreased. Conclusion Knockdown of TAZ can affect the proliferation and apoptosis of TSCC cell line CAL27 by modulating the expression of proteins related to cell cycle and apoptosis.

Key words: TAZ gene, Tongue squamous cell carcinoma CAL27 cell line, Cell proliferation, Cell apoptosis

中图分类号:

R739.8

陈希彦,王琪,顾伟亭,文勇. 干扰TAZ基因对舌鳞癌细胞CAL27增殖凋亡的影响及其机制[J]. 山东大学学报 (医学版), 2018, 56(10): 79-85.

CHEN Xiyan, WANG Qi, GU Weiting, WEN Yong. Effects of TAZ knockdown on the proliferation and apoptosis of TSCC cell line CAL27 and the molecular mechanism[J]. Journal of Shandong University (Health Sciences), 2018, 56(10): 79-85.

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