过表达MIPU1降低阿霉素所致人胚胎肾HEK293细胞的凋亡

Overexpression of MIPU1 reduces doxorubicin-induced apoptosis in human embryonic kidney 293 cells

目的探讨过表达MIPU1对阿霉素(Doxorubicin,DOX)所致细胞凋亡的影响及其可能的分子机制,为预防DOX抗肿瘤治疗过程中的毒副作用提供新的思路。方法 1)MTT实验观察DOX处理下过表达MIPU1对细胞存活率的影响;2)Hoechst染色和流式细胞术分析过表达MIPU1对DOX所致细胞凋亡的影响;3)RT-PCR和Western blot分析过表达MIPU1对DOX所致Bcl-2、P53和Bax在mRNA水平和蛋白水平上表达变化的影响。结果过表达MIPU1后,细胞存活率明显升高(P<0.05 vs p EGFP+DOX),凋亡率显著下降(P<0.01 vs p EGFP+DOX),P53和Bax的表达显著下降(P<0.05 vs p EGFP+DOX),而Bcl-2的表达则明显增多(P<0.05 vs p EGFP+DOX)。结论过表达MIPU1能降低DOX引起的HEK293细胞凋亡,可能与抑制P53和Bax的表达同时促进Bcl-2的表达有关。

Objective To explore the affection of overexpression MIPU1 on DOX-mediated apoptosis and the mechanisms,so to provide novel insight into the biological functions of Mipu1,and a new clue for preventing the side-effects of DOX during anticancer implication.Methods Apoptosis was induced by Doxorubicin in this experiment.The eukaryotic expression plasmid pEGFP-MIPU1 was constructed previously and delivered into the cells to increase the expression of MIPU1.Firstly,MTT assay was used to estimate the cells viability.Then apoptosis was assessed by Hoechst staining and flow cytometry （FCM）.Thirdly,Western blot and RT-PCR were performed to detect the expression of Bax,P53 and Bcl-2 at the mRNA and protein levels respectively.Results After transfected with pEGFP-MIPU1,the overexpression of MIPU1 significantly attenuated the mortality rate induced by DOX （P ＜ 0.05 vs pEGFP ＋ DOX）,and markedly decreased the cellular apoptosis treated with DOX （P ＜0.01 vs pEGFP ＋ DOX）; The overexpression MIPU1 inhibited the increase of P53 and Bax induced by DOX （P ＜ 0.05 vs pEGFP ＋ DOX）,while reversed the decrease of Bcl-2 expression in DOX-induced HEK293 cells （P ＜ 0.05 vs pEGFP ＋ DOX）.Conclusions Overexpression MIPU1 inhibites apoptosis in HEK293 cells induced by DOX,and MIPU1 may decrease the expression of P53 and Bax,and promote the expression of Bcl-2 in DOX-induced HEK293 cells.