过表达LATS2基因诱导口腔鳞癌细胞自噬的初步研究

The effect of overexpressed LATS2 gene on the autophagy of oral squamous cell carcinoma cells

目的:探讨大肿瘤抑制因子2(LATS2)基因过表达诱导口腔鳞癌细胞(OSCC)发生自噬性的现象的机制。方法:构建慢病毒载体LATS2稳定转染SCC-25细胞,并设空载体为对照组。转染72 h后提取细胞总蛋白,Western印迹方法检测LATS2、自噬标志物(LC3)、Beclin-1蛋白表达的变化,双荧光自噬流以及透射电子电镜检测细胞自噬水平。结果:与对照组相比,实验组细胞LC3、Beclin-1蛋白的表达明显增多(P<0.05);透射电子显微镜观察到OSCC细胞SCC-25细胞质内有大量自噬体和自噬溶酶体。结论:LATS2基因可诱导OSCC细胞SCC-25发生自噬。

Objective:To determine the autophagy of oral squamous cell carcinoma(OSCC)cells induced by large tumor suppressor homolog 2(LATS2)gene overexpression,and to investigate its potential mechanism.Methods:OSCC SCC-25 cells were transfected with Lentivirous-LATS2,and the empty vector was used in control group.At 72 h after the transfection,the total protein were extracted.The changes of LATS2 and the expression of autophagy marker(LC3 and Beclin-1)proteins was determined by Western blotting.The mRFP-GFP-LC3 label and transmission electron microscopy(TEM)were used to determine the autophagy level and ultrastructure.Results:Compared with the control group,the expression of LC3 and Beclin-1 in the experimental cells was significantly increased(P<0.05),mRFP-GFP-LC3 label and TEM showed that autophagosomes and autolysosomes were found significantly increased in the cytoplasm of SCC-25 cells.Conclusion:LATS2 gene can induce autophagy in OSCC cells.