The effect of silencing human telomerase reverse transcriptase by RNA interference on U87 glioma cell proliferation and apoptosis

Abstract

Objective To observe the effect of human telomerase reverse transcriptase (hTERT) on U87 glioma cell proliferation and apoptosis.Methods The small interfering RNA (siRNA) expression plasmid targeting hTERT was constructed and transfected into U87 glioma cells.The expression level of hTERT protein and mRNA was detected by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR) respectively.The U87 glioma cell proliferation and apoptosis were analyzed by methyl thiazol tetrazolium (MTT) assay and flow cytometry,respectively.Results Compared with the blank control group and the negative control group,the expression level of hTERT protein in each siRNA group was significantly reduced (P ＜ 0.05).The relative expression level of hTERT mRNA in each siRNA group was 0.79 ±0.11,0.58 ±0.15 and 0.39 ±0.16 respectively,which was significantly lower than in the negative control group (P ＜ 0.05).As compared with the blank control group and the negative control group,the proliferation of U87 glioma cells after transfection was significantly inhibited and the strongest inhibitory effect was observed on the third day.The apoptosis rate of U87 glioma cells of in each siRNA group which was transfected after 48 h was (14.31 ± 0.93) ％,(17.57 ± 0.61) ％ and (22.68 ± 1.04) ％ respectively,which was significantly higher than in the blank control group and the negative control group (P ＜0.05).Conclusion The silence of hTERT gene after siRNA targeting hTERT being transfected into U87 glioma cells can significantly inhibit cell proliferation and induce cell apoptosis. Key words: RNA interference; Human telomerase reverse transcriptase; Glioma; Cell proliferation