显性负突变hTERT在TSA诱导Hela细胞凋亡中的作用

Effect of dominant negative human telomerase reverse transcriptase on apoptosis of Hela induced by trichostatin A

目的通过转染显性负突变hTERT探讨其在TSA诱导宫颈癌细胞株Hela凋亡中的作用。方法PCR-TRAP-ELISA法检测转染显性负突变,野生型hTERT和对照质粒以后Hela细胞的端粒酶活性。磺酰罗丹明B(sulforhodamine B,SRB)法检测细胞生长率;AnnexinⅤ/PI法检测各组转染细胞在TSA作用下的早期凋亡。结果转染显性负突变hTERT的Hela细胞的端粒酶活性显著低于对照组,2μmol/LTSA作用对照细胞Hela-puro、Hela-DNhTERT和Hela-WThTERT细胞,Hela-DNhTERT相对于Hela-puro生长率差异明显,Hela-DNhTERT细胞凋亡率明显高于Hela-WThTERT和Hela-puro组。结论转染DNhTERT对TSA诱导的Hela细胞凋亡具有协同作用。

[Objiective] This study was designed to investigate effect of dominant negative human telomerase reverse transeriptase on apoptosis of Hela induced by trichostatin A, [Methods] the telomerase activity was detected by PCR and telomeric repeat amplification protocol assay （PCR-TRAP-ELISA）. Sulforhodamine B method was employed to determine the growth rate of Hela cells at different time. The early apoptosis was measured by Annexin V/ PI stain and flow cytometry, [Results] After transfected the dominant negative, wild type hTERT and control plasmid into Hela cells, the significant difference of telomeras activity of Hela cells was observed by PCR-TRAP-ELISA assay. DN-hTERT transfected HeLa cells with less telomeras activity were more susceptible to apoptosis induced by trichostatin A. [Conclusion] DN-hTERT transfection might have co-effection on apoptosis of Hela cells induced by trichostatin A.