PTEN重组腺病毒对胶质瘤细胞株凋亡及凋亡抑制蛋白因子家族的影响

Effects of PTEN overexpression on apoptosis and IAPs in glioma cell lines

目的探讨以腺病毒为载体转染第10号染色体缺失的磷酸酶和张力蛋白同源基因(phosphatase and tensin homology deleted on chromosome ten,PTEN)对胶质瘤细胞株增殖和凋亡的影响,并探讨其相关作用机制。方法以携带PTEN的重组腺病毒(Ad-PTEN)转染胶质瘤细胞株,应用RT-PCR、Western blot检测PTEN基因mR-NA、蛋白表达的变化,MTT、TUNEL和流式细胞法检测转染后细胞增殖能力及凋亡率的改变。RT-PCR检测转染后凋亡抑制蛋白因子家族(inhibtor of apoptosis family of proteins,IAPS):CIAP1、CIAP2、XIAP、SURVIVIN的mRNA水平的变化,Westernblot检测细胞CIAP1、CIAP2、XIAP、SURVIVIN的蛋白表达水平的变化。结果经荧光显微镜证实Ad-PTEN转染成功,Western blot、RT-PCR显示转染后PTEN基因表达显著增加。PTEN转染U251、U87、A172细胞株后3d细胞凋亡率明显高于对照组(F分别为93.80、95.70、72.52,P<0.01),PTEN转染组中CIAP1、CIAP2、XI-AP、SURVIVIN mRNA及蛋白水平与空白转染组mRNA及蛋白水平相比明显下降(F分别为73.8、49.5、50.4、69.1、39.7、48.8、54.3、70.3,P<0.01)。结论 PTEN转染后可能通过抑制IAPS家族表达途径促进细胞凋亡,从而抑制胶质瘤细胞株的的生长。

Objective To investigate the effects of overexpression of phosphatase and tensin homology deleted on chromosome ten（PTEN） on apoptosis in glioma cell lines.Methods The mRNA and protein expression levels of PTEN in glioma cell lines transfected by Ad-PTEN were determined using RT-PCR and Western blotting.Apoptotic cell death was detected using MTT、TUNEL and Flow cytometry.The mRNA and protein expression levels of IAPs were determined using RT-PCR and Western blotting.Results The Ad-PTEN transfection was confirmed using fluorescence microscopy.Western blot and RT-PCR showed that apoptosis were significantly higher and the mRNA and protein expression levels of IAPs in glioma cells were significantly lower in glioma cells transfected by Ad-PTEN.Conclusions Overexpression of PTEN inhibits glioma cell growth through inducing cell apoptosis and inhibiting the expression of IAPs.