Cyclopamine对前列腺癌LNCaP细胞增殖凋亡及PSAmRNA基因表达的影响  

Effects of Cyclopamine on proliferation,apoptosis and expression of PSAmRNA in prostate cancer LNCaP cells

作　　者：吕志勇[1] 吕凌东 吴小侯[2] 

机构地区：[1]宁夏医科大学附属医院泌尿外科,银川750004 [2]重庆医科大学附属第一医院泌尿外科,400016

出　　处：《重庆医学》2014年第27期3584-3587,共4页Chongqing medicine

基　　金：宁夏回族自治区卫生厅重点科研计划项目支助

摘　　要：目的探讨Cyclopamine对前列腺癌LNCaP细胞增殖、凋亡的作用及对PSAmRNA基因表达的影响。方法不同水平Cyclopamine(1、5、10、15μmol/L)干预LNCaP细胞不同时间(24、48、72h),MTT法检测其对细胞增殖的抑制,Hoechst33258染色观察细胞形态变化,流式细胞术检测细胞凋亡率,实时荧光定量逆转录聚合酶链反应(FQ-RT-PCR)观察PSAmRNA基因表达的影响。结果 5、10、15μmol/L组Cyclopamine对LNCaP细胞增殖有显著抑制作用,与对照组比较差异有统计学意义(P<0.01),10μmol/L组于48h达到IC50;10、15μmol/L组的24、48、72h凋亡率分别为37.21%、57.38%、57.98%和21.16%、71.31%、72.90%,与对照组比较差异有统计学意义(P<0.01)。细胞凋亡形态的改变随着Cyclopamine水平增大和作用时间的延长也显著增强。PSAmRNA基因的表达水平随着水平的上升呈现明显的递减趋势,较对照组显著降低(P<0.01);10、15μmol/L水平的Cyclopamine在不同时间段PSAmRNA基因的表达均极低。结论 Cyclopamine能够明显抑制LNCaP细胞的增殖,诱导细胞凋亡,并显著下调LNCaP细胞PSAmRNA基因的表达,一定水平Cyclopamine可能对晚期前列腺癌的治疗有效。Objective To investigate the effects of cyclopamine on the proliferation ,apoptosis and the expression of PSAmRNA in prostate cancer LNCaP cells .Methods LNCaP cells were interfered with different concentrations of cyclopamine （1 ,5 ,10 ,15μmol/L） at the different timepoints（24 ,48 ,72 h） .The poliferation inhibition was measured by the MTT assay ;the apoptotic mor-phological changes were observed by the Hoechst33258 staining method ;the apoptosis rate was examined by the flow cytometry ;the effects of PSAmRNA gene expression was detected by the FQ-RT-PCR .Results 5 ,10 ,15μmol/L cyclopamine groups had obvious inhibition effect on the LNCaP cell proliferation ,which had statistically significant difference compared with the control group （P〈0 .01） .10 μmol/L group reached IC50 at 48 h ;the apoptosis rates at 24 ,48 ,72 h in the 10 ,15 μmol/L groups were 37 .21% , 57 .38% ,57 .98% and 21 .16% ,71 .31% ,72 .90% respectively ,the difference had statistically significant difference compared with the control group（P〈0 .01） .The cellular apoptotic morphological changes were significantly enhanced with the increase of cyclo-pamine concentration and the extension of action time .The expression level PSAmRNA gene exhibited the obvious decreasing trend with the increase of cyclopamine concentration and was significantly decreased compared with the control group （P〈 0 .01） .The PSAmRNA gene expression was extremely low in 10 ,15 μmol/L cyclopamine at different time period .Conclusion Cyclopamine significantly inhibits the proliferation of LNCaP cells ,induces apoptosis and obviously down-regulates the PSAmRNA gene expres-sion in LNCaP cells .Certain concentration of cyclopamine may be effective for treating advanced prostate cancer .

关 键 词：前列腺肿瘤 细胞增殖 

分 类 号：R737.25[医药卫生—肿瘤]