论文部分内容阅读
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。
AIM: To investigate the effect of down-regulation of Pim-3 on the apoptosis of tongue squamous cell carcinoma cells and to explore the molecular mechanism of apoptosis. METHODS: Pim-3 siRNA and control siRNA were transfected into Tca8113 tongue squamous cell carcinoma respectively, and another untreated group was established. The expression of Pim-3 mRNA and protein in the three groups of cells before and after transfection was analyzed by real-time fluorescence quantitative PCR and Western blotting. The changes of cell proliferation were analyzed by CCK-8 reagent before and after transfection, and the expression of Pim-3 was detected by FCM Effect of down regulation on apoptosis of tongue squamous cell carcinoma Tca8113 cells. Finally, the changes of the expression of apoptosis-related proteins Bcl-2, Bax, Bad, pBad112, pBad136 and pBad155 before and after transfection were analyzed by Western blotting. Results: Pim-3 siRNA could effectively down-regulate the expression of Pim-3 mRNA and protein in tongue squamous cell carcinoma Tca8113 cells. The down-regulation of Pim-3 expression can significantly inhibit the proliferation of Tca8113 cells and induce their apoptosis. In addition, down-regulation of Pim-3 significantly up-regulated the expression of Bax protein and down-regulated the expression of Bcl-2 and pBadl12 (P <0.05), but did not change the expression of total Bad protein and other Bad phosphorylated forms (pBad136 and pBad155) . CONCLUSIONS: Pim-3-induced down-regulation of apoptosis may be closely related to the increase of Bax expression and the decrease of Bcl-2 and pBad112 expression. Therefore, Pim-3 is expected to be a molecular target for the treatment of tongue squamous cell carcinoma.