Pim-3表达下调对舌鳞状细胞癌细胞凋亡的影响及其分子机制研究  

Effect of down-regulation of Pim-3 expression on apoptosis of tongue squamous-cell carcinoma and its molecular mechanism

作　　者：焦静[1] 刘红涛[2] 李沙[3] 

机构地区：[1]郑州人民医院口腔科,郑州450003 [2]郑州大学生物工程系,郑州450001 [3]郑州大学第一附属医院口腔科,郑州450052

出　　处：《肿瘤》2011年第6期489-494,共6页Tumor

摘　　要：目的:研究Pim-3表达下调对舌鳞状细胞癌细胞凋亡的影响,并探讨其凋亡的相关分子机制。方法:将Pim-3siRNA和对照siRNA分别转染舌鳞状细胞癌Tca8113细胞,另设未处理组。利用实时荧光定量PCR和蛋白质印迹法分析转染前后3组细胞中Pim-3mRNA和蛋白的表达,利用CCK-8试剂分析转染前后细胞增殖能力的变化,并进一步采用FCM法检测Pim-3表达下调对舌鳞状细胞癌Tca8113细胞凋亡的影响。最后,利用蛋白质印迹法分析转染前后细胞凋亡相关蛋白Bcl-2、Bax、Bad、pBad112、pBad136和pBad155表达的变化。结果:Pim-3siRNA能有效下调舌鳞状细胞癌Tca8113细胞中Pim-3mRNA和蛋白的表达。Pim-3表达的下调能明显抑制Tca8113细胞的增殖并诱导其凋亡。此外,Pim-3表达下调能显著上调Bax蛋白的表达和下调Bcl-2和pBad112的表达(P<0.05),但不改变总的Bad蛋白和其他Bad磷酸化形式(pBad136和pBad155)蛋白的表达。结论:Pim-3表达下调诱导的细胞凋亡可能与Bax表达水平上升及Bcl-2和pBad112表达水平下降密切相关,因而Pim-3有望成为舌鳞状细胞癌治疗的分子靶点。Objective：To investigate the effect of down-regulation of Pim-3 expression mediated by RNA interference on apoptosis of tongue squamous-cell carcinoma,and to explore its molecular mechanism.Methods：Pim-3 siRNA and single siRNA were transfected into tongue squamous-cell carcinoma Tca8113 cells,respectively.The expressions of Pim-3 mRNA and protein were detected by real-time fluorescence quantitative PCR and Western blotting.The change of cell proliferation capability was analyzed using CCK-8 kit.The effect of down-regulation of Pim-3 expression on apoptosis of Tca8113 cells was examined by flow cytometry.Finally,the expression changes of apoptosis-related proteins such as Bcl-2,Bax,Bad,pBad^112,pBad^136 and pBad^155 were investigated by Western blotting.Results：Pim-3 siRNA could effectively down-regulate the expressions of Pim-3 mRNA and protein in Tca8113 cells.The down-regulation of Pim-3 expression obviously inhibited the cell proliferation and induced the apoptosis of Tca8113 cells.Furthermore,the down-regulation of Pim-3 expression could significantly elevate the expression of Bax protein and decrease the expressions of Bcl-2 and pBad^112 proteins（P〈0.05）,but the expression levels of total Bad protein and the phosphorylated types of Bad（pBad^136 and pBad^155） were not changed.Conclusion：The apoptosis induced by down-regulation of Pim-3 expression may be tightly associated with the increase of Bax expression and decrease of Bcl-2 and pBad^112 expressions.Pim-3 may become a molecular target in the therapy for tongue squamous-cell carcinoma.

关 键 词：舌肿瘤 基因表达调控 肿瘤 RNA干扰 原癌基因蛋白质pim-3 细胞凋亡 凋亡调控蛋白类 

分 类 号：R739.86[医药卫生—肿瘤]