PTEN基因调控口腔鳞癌细胞TCA-8113增殖及凋亡  

PTEN gene expression inhibits proliferation and promotes apoptosis in human oral squamous carcinoma cell line TCA-8113

作　　者：杨雨晗[1] 邓缅[2] 

机构地区：[1]成都医学院生物医学系细胞生物学和遗传学教研室,成都610083 [2]成都医学院生物医学系生物化学与分子生物学教研室,成都610083

出　　处：《第三军医大学学报》2011年第14期1484-1487,共4页Journal of Third Military Medical University

基　　金：四川省教育厅科研基金(10ZC090)~~

摘　　要：目的评估PTEN基因在人口腔鳞癌细胞系TCA-8113中的增殖抑制和促凋亡功能。方法利用PTEN真核表达质粒转染TCA-8113细胞,另设PBS处理作为空白组,转染pcDNA3.1质粒和Lipofectamine处理作为对照组。采用MTT和克隆形成实验检测细胞增殖情况,采用流式细胞术、TUNEL和Western blot法检测细胞凋亡。结果 pcDNA3.1-PTEN转染TCA-8113细胞得到高效的外源性表达。克隆形成实验结果表明,空白组、Lipofectamine处理组、空载体质粒组和PTEN转染组的克隆数分别为(107.34±4.62)、(79.27±3.71)、(72.03±2.38)和(19.73±1.02),PTEN转染组抑制率为72.60%,与空载体质粒组相比,差异有统计学意义(P<0.05);MTT实验结果显示PTEN基因的表达对TCA-8113细胞的抑制率在48 h和72 h分别为18.16%和42.74%,PTEN转染组与空载体质粒组比较有统计学差异(P<0.05);流式细胞术检测结果显示,PTEN基因能引起62.12%的细胞死亡;TUNEL检测PTEN基因高表达能引起(45.67±2.31)%的细胞凋亡;Western blot检测结果表明,PTEN能抑制Bcl-2的表达并激活Bax的表达。结论 PTEN在TCA-8113细胞中的表达能显著抑制细胞增殖,促进细胞凋亡。Objective To evaluate the effects of PTEN gene expression on inhibiting the proliferation and promoting the apoptosis of human oral squamous carcinoma cell line TCA-8113.Methods TCA-8113 cells were transfected with eukaryotic expression plasmid pcDNA3.1-PTEN to obtain an experimental group.Besides,TCA-8113 cells were also treated with PBS,plasmid pcDNA3.1 and Lipofectamine to obtain a blank group and two control groups（a pcDNA3.1-transfected group and a Lipofectamine-treated group）.The proliferation of TCA-8113 cells was detected by MTT assay and colony-forming assay.The apoptosis of TCA-8113 cells was evaluated by flow cytometry,TUNEL assay and Western blotting.Results TCA-8113 cells transfected with plasmid pcDNA3.1-PTEN had high intracellular expression of PTEN.Colony-forming assay showed that the clone numbers of the blank group,Lipofectamine-treated group,pcDNA3.1-transfected group and pcDNA3.1-PTEN-transfected group were 107.34±4.62,79.27±3.71,72.03±2.38 and 19.73±1.02,respectively.The inhibitive rate of TCA-8113 cells in the pcDNA3.1-PTEN-transfected group was 72.60%,significantly higher than that in the pcDNA3.1-transfected group（P0.05）.MTT assay showed that the inhibitive rates of TCA-8113 cells in the pcDNA3.1-PTEN-transfected group were 18.16% and 42.74% at 48 h and 72 h after transfection,significantly higher than those in the pcDNA3.1-transfected group（P0.05）.Flow cytometry showed that the death rate of TCA-8113 cells in the pcDNA3.1-PTEN-transfected group was 62.12%,significantly higher than that in the pcDNA3.1-transfected group（P0.05）.TUNEL assay showed that the apoptosis of TCA-8113 cells in the pcDNA3.1-PTEN-transfected group was（45.67±2.31）%,significantly higher than that in the pcDNA3.1-transfected group（P0.05）.Furthermore,PTEN inhibited Bcl-2 expression and activated Bax expression,as proven by Western blotting.Conclusion The expression of PTEN gene can remarkably inhibit the proliferation and promote the apoptosis of TCA-8113 cells.

关 键 词：口腔鳞癌 PTEN 增殖 凋亡 

分 类 号：R739.85[医药卫生—肿瘤] R394.2[医药卫生—临床医学]