摘要
目的探讨PTEN对食管癌细胞增殖的影响及作用机制。方法将食管鳞状细胞癌细胞株TE1和TE13分为对照组、空载体组和转染组3个处理组。对照组仅常规培养细胞,而空载体组和转染组常规培养细胞后,分别转染空载体质粒、PTEN/sh-AKT质粒后备用。采用细胞计数法检测细胞浓度,MTT方法检测细胞增殖能力,RT-PCR方法检测细胞PTEN、AKT基因的表达,Western blot方法检测细胞PTEN、p-AKT蛋白的表达。结果转染PTEN质粒后,转染组两种细胞浓度均较其余两组明显下降(P<0.05)。转染PTEN质粒后,转染组两种细胞的增殖能力(OD值)均较其余两组明显下降(P<0.05)。转染PTEN质粒后,转染组两种细胞PTEN基因表达量均明显高于其余两组(P<0.05),而AKT基因无明显变化(P>0.05);转染sh-AKT质粒后,各组间两种细胞PTEN、AKT基因表达量差异均无统计学意义(P>0.05)。转染PTEN质粒后,转染组两种细胞PTEN蛋白表达量均明显高于其余两组(P<0.05),而p-AKT蛋白表达量均明显低于其余两组(P<0.05);转染sh-AKT质粒后,各组间两种细胞PTEN蛋白表达量差异均无统计学意义(P>0.05),而转染组两种细胞p-AKT蛋白表达量均明显低于其余两组(P<0.05)。结论基因水平上调PTEN的表达可抑制AKT的活化,进而达到抑制食管癌细胞增殖的作用。
Objective To investigate the effects of PTEN on esophageal carcinoma cell proliferation and its mech-anism. Methods Esophageal squamous cancer cell TEl and TEl3 were divided into three groups, including controlgroup, empty plasmid transfected group and plasmid transfected group. Cells in control group were cultured in convention-al condition. Empty plasmid and plasmid PTEN/sh - AKT were amplified and transfected into the cells in empty plasmidtransfected group and plasmid transfeeted group, respectively, after conventional culture. Cell concentration was examinedin cell count method, and cell proliferation ability was examined by MTF. PTEN and AKT genes expressions were detectedby RT - PCR, and PTEN and p - AKT protein expressions were examined by Western blot. Results .After transfection ofPTEN, the cell concentrations of TEl and TEl3 were significantly reduced in plasmid transfected group than those in theother two groups （ P 〈 0. 05 ） ; so were the cell proliferation abilities （01） value ） in plasmid transfected group （ P 〈0. 05 ）. After transfection, PTEN gene expression was significantly up - regulated in plasmid transfected group than thosein the other two groups （ P 〈 0.05 ）, while no significant difference in AKT was observed （ P 〉 0. 05 ）. After transfectionof plasmid sh - AKT, no significant difference in PTEN or AKT gene expression was observed among the three groups （ P 〉 0. 05 ）. After transfection of plasmid PTEN, PTEN protein expression was significantly up - regulated in plasmid trans-fected group than those in the other two groups （ P 〈 0. 05 ） , however, significant down - regulation in p - AKT observed（ P 〈 0.05 ）. After transfection of plasmid sh - AKT, there was no significant difference in PTEN protein expression amongthe three groups, but significant down - regulation of p - AKT in plasmid transfected group was observed （ P 〈 0. 05 ）.Conclusion PTEN may inhibit the activation of AKT, and then suppresses the proliferation ability of esoph
出处
《广东医学》
CAS
CSCD
北大核心
2014年第24期3773-3777,共5页
Guangdong Medical Journal
基金
国家自然科学基金资助项目(编号:81303271)
河北省医学科学研究重点课题计划(编号:20130545)
