Survivin调控信号通路PTEN/PI3K/AKT对CNE-2细胞增殖和凋亡机制研究

Mechanism study on survivin regulated the signaling pathway PTEN/PI3K/AKT in proliferation and apoptosis of CNE-2 cells

目的检测鼻咽癌CNE-2细胞中Survivin表达变化与信号通路PTEN/PI3K/AKT的表达差异,探讨对鼻咽癌CNE-2细胞的增殖和凋亡产生的影响。方法设计特异性siRNA转染CNE-2细胞,抑制细胞内Survivin表达;构建过表达质粒转染CNE-2细胞,促进细胞内Survivin表达;以CCK8法检测各组中不同时间CNE-2细胞活性; RT-PCR法检测CNE-2细胞中Survivin、PI3K、PTEN、AKT的mRNA表达。蛋白免疫印迹法检测CNE-2细胞中Survivin、PI3K、PTEN、AKT、p-AKT各相关蛋白表达情况。结果 (1)CCK8:利用CCK8检测细胞活性,与空白对照组相比,Survivin抑制组活性降低; Survivin过表达组活性增加。(2) RT-PCR结果显示:Survivin抑制组Survivin mRNA表达抑制率为(41. 9±6. 9)%(P <0. 01),PI3K mRNA表达抑制率为(43. 3±7. 2)%(P <0. 01),AKT mRNA表达抑制率为(38. 7±4. 1)%(P <0. 01),PTEN mRNA表达抑制率无统计学意义; Survivin过表达组Survivin mRNA过表达率为(203. 8±17. 1)%(P <0. 01),AKT mRNA过表达率为(239. 0±34. 2)%(P <0. 01),PTEN、PI3K mRNA表达率无统计学意义。(3) Western blot结果:Survivin抑制组与阴性对照组相比,PTEN、p-AKT蛋白表达量无明显差异,Survivin、PIK3、AKT蛋白表达量显著下调。Survivin过表达组与阴性对照组相比,PTEN、PIK3、p-AKT蛋白表达量无明显差异,Survivin、AKT蛋白表达量显著上调。结论在鼻咽癌CNE-2细胞中,Survivin调控AKT,与AKT呈正相关。Survivin抑制时,能有效抑制CNE-2细胞的增殖,诱导细胞发生凋亡; Survivin过表达时,能有效促进CNE-2细胞增殖,抑制其发生凋亡。对细胞的增殖及凋亡起正协同作用。

Objective To investigate the Survivin expression and PTEN/PI3 K/AKT pathway expression in nasopharyngeal carcinoma( NPC) CNE-2 cells,and to explore its effect on the proliferation and apoptosis of NPC. Methods The specific siRNA was designed to transfect CNE-2 cells to inhibit the expression of Survivin,while the expression of Survivin in CNE-2 cells was enhanced by the construction of expression plasmids. The activity of CNE-2 cells at different times was detected by CCK8 assay. The nRNA and protein levels of Survivin,PI3 K,PTEN and AKT mRNA were assessed by RT-PCR assay and Western blot,respectively. Results Compared with the control group,the activity of Survivin inhibited group was significantly lower,while the activity of Survivin over-expression group was significantly increased. The results of RT-PCR showed that the inhibition rates of Survivin mRNA,PI3 K mRNA and AKT mRNA were( 41. 9 ± 6. 9)%,( 43. 3 ± 7. 2)% and( 38. 7 ± 4. 1)%,respectively( P < 0. 01). The overexpression rates of Survivin mRNA and AKT mRNA were( 203. 8 ± 17. 1)% and( 239. 0 ± 34. 2)% in Survivin overexpression group( P < 0. 01),while there was no significant difference in the relative expression of PI3 K or PTEN mRNA( P > 0. 05). There was no significant difference in the expression of PTEN or p-AKT protein between the Survivin inhibition group and the negative control group. The expression of AKT protein was significantly down-regulated,while the expression of PTEN,PI3 K and p-AKT protein was significantly up-regulated in the Survivin overexpression group compared with the negative control group. Conclusion In CNE-2 cells,the regulation of AKT by Survivin is positively correlated with AKT. When Survivin is inhibited,it can effectively inhibit the proliferation and induce the apoptosis of CNE-2 cells. Survivin overexpression can effectively promote the proliferation of CNE-2 cells and inhibit their apoptosis. It has positive synergistic effect on cell proliferation and apoptosis.