敲低富含亮氨酸的三角状五肽重复结构蛋白(LRPPRC)促进激素抵抗性前列腺癌细胞凋亡

Knock-down of leucine-rich pentatricopeptide repeat containing (LRPPRC) promotes apoptosis of hormone resistant prostate cancer cells

目的研究富含亮氨酸的三角状五肽重复结构蛋白(LRPPRC)对激素抵抗性前列腺癌细胞凋亡的影响。方法Western blot法检测前列腺癌DU145细胞和LNCaP细胞LRPPRC和雄激素受体(AR)蛋白水平;在DU145细胞中瞬转LRPPRC小干涉RNA(siLRPPRC);反转录PCR检测转染细胞LRPPRC mRNA水平;Western blot法检测LRPPRC蛋白水平;噻唑蓝(MTT)法检测细胞存活率;Hoechst33258法和流式细胞术检测细胞凋亡率;荧光素酶法检测细胞ATP水平;Western blot法检测Bcl2、BAX和胱天蛋白酶3(caspase-3)蛋白水平。结果 LRPPRC在DU145细胞和LNCaP细胞均高表达,但DU145细胞不表达AR。敲低DU145细胞LRPPRC水平后,细胞存活率显著降低、细胞凋亡增加、 ATP水平下降、活化的caspase-3蛋白水平增高、Bcl2蛋白水平降低。结论敲低激素抵抗性前列腺癌DU145细胞LRPPRC促进细胞凋亡。

Objective To investigate the effects of leucine-rich pentatricopeptide repeat containing(LRPPRC) on proliferation and apoptosis in hormone resistant prostate cancer cells. Methods LRPPRC and androgen receptor(AR) expressions in DU145 and LNCaP cells were detected by Western blot analysis. The siRNA targeting LRPPRC gene was transiently transfected into DU145 cells. LRPPRC expression was detected by reverse transcription PCR and Western blot analysis. The cell survival rate was analyzed by MTT assay. ATP was measured by luciferin-luciferase assay. The cell apoptosis was investigated by flow cytometry and Hoechst 33258 staining. Caspase-3, Bcl2 and BAX protein expressions were detected by Western blot analysis. Results DU145 and LNCaP cells highly expressed LRPPRC. LNCaP cells highly expressed AR, while no expression of AR was observed in DU145 cells. Down-regulation of LRPPRC decreased cell survival rate and ATP level, increased apoptosis rate and caspase-3 activation, and decreased Bcl2 expression in DU145 cells. Conclusion Knock-down of LRPPRC promotes apoptosis of hormone resistant prostate cancer cells.