CAJ | 학술논문

目的观察雷公藤内酯醇( triptolide,TPL)对去势后抵抗性前列腺癌(Castration-Resistant Prostate Cancer,CRPC) PC3细胞的生长增殖抑制作用,对其诱导凋亡相关基因进行分析.方法 MTT比色法观察TPL对PC3细胞的生长增殖抑制作用；Annexin-V/PI标记分析细胞凋亡;RT-PCR方法检测0、6、12、24 h BCL-2、BAX、PIG3、P21、FAS、CASPASE3等与凋亡相关基因表达变化.结果 TPL抑制PC3细胞生长呈时间和剂量依赖性,24h和48 h半数抑制浓度分别为18.3 ng/ml和13.5ng/ml,与24h组相比,48 h组低浓度(5 ng/ml,t=1.47,P＞0.05)和高浓度(160 ng/ml,t =0.91,P＞0.05)差异无统计学意义.而10 ng/ml(t=3.26,P＜0.05)、20 ng/ml(t=4.21,P＜0.05)、40ng/ml(t=4.09,P＜0.05)、80ng/ml(t=2.91,P＜0.05)差异有统计学意义.Annexin-V/PI检测经18.3 ng/ml TPL处理后的PC3细胞凋亡随着作用时间的延长而增多,相对于对照组,6、12、24 h组Anexin-V阳性/PI阴性表达率分别为(5.42±2.21)％(t=3.52,P＜0.05)、(13.51±3.37)％(t=6.53,P＜0.01)、(29.3±4.53)％(t=8.74,P＜0.01)差异有统计学意义,并且各组间差异有统计学意义(P＜0.05)；RT-PCR显示经18.3 ng/ml TPL处理后,BAX、PIG3表达递增,P21、BCL-2表达递降,FAS、CASPASE3表达无明显改变.结论 TPL可以抑制PC3生长增殖并诱导细胞凋亡,而在凋亡的过程中,BAX、BCL-2、PIG3、P21等基因的改变可能扮演着重要的角色.
목적 관찰뢰공등내지순( triptolide,TPL)대거세후저항성전렬선암(Castration-Resistant Prostate Cancer,CRPC) PC3세포적생장증식억제작용,대기유도조망상관기인진행분석.방법 MTT비색법관찰TPL대PC3세포적생장증식억제작용；Annexin-V/PI표기분석세포조망;RT-PCR방법검측0、6、12、24 h BCL-2、BAX、PIG3、P21、FAS、CASPASE3등여조망상관기인표체변화.결과 TPL억제PC3세포생장정시간화제량의뢰성,24h화48 h반수억제농도분별위18.3 ng/ml화13.5ng/ml,여24h조상비,48 h조저농도(5 ng/ml,t=1.47,P＞0.05)화고농도(160 ng/ml,t =0.91,P＞0.05)차이무통계학의의.이10 ng/ml(t=3.26,P＜0.05)、20 ng/ml(t=4.21,P＜0.05)、40ng/ml(t=4.09,P＜0.05)、80ng/ml(t=2.91,P＜0.05)차이유통계학의의.Annexin-V/PI검측경18.3 ng/ml TPL처리후적PC3세포조망수착작용시간적연장이증다,상대우대조조,6、12、24 h조Anexin-V양성/PI음성표체솔분별위(5.42±2.21)％(t=3.52,P＜0.05)、(13.51±3.37)％(t=6.53,P＜0.01)、(29.3±4.53)％(t=8.74,P＜0.01)차이유통계학의의,병차각조간차이유통계학의의(P＜0.05)；RT-PCR현시경18.3 ng/ml TPL처리후,BAX、PIG3표체체증,P21、BCL-2표체체강,FAS、CASPASE3표체무명현개변.결론 TPL가이억제PC3생장증식병유도세포조망,이재조망적과정중,BAX、BCL-2、PIG3、P21등기인적개변가능분연착중요적각색.
Objective To detect the treatment effect of PC3 cells with triptolide on altering the expression of genes.Methods MTT assay was used to detec the inhibition of proliferation.Apoptosis was detected by Annexin-V/PI staining.RT-PCR was used to analyze the mRNA expressions of BCL-2,BAX,PIG3,P21,FAS,CASPASE3.Results Triptolide caused a time - and dose - dependent inhibition of cell proliferation,and IC50 of 24 h and 48 h were 18.3 ng/ml and 13.5 ng/ml,respectively.Compared with the 24 h group,the low concentration of triptolide(5 ng/ml,t =1.47,P ＞0.05)and the high concentration of triptolide ( 160 ng/ml,t =0.91,P ＞0.05)had no statistical significance in 48 h group,while 10 ng/ml( t =3.26,P ＜0.05),20 ng/ml( t =4.21,P ＜0.05),40 ng/ml( t =4.09,P ＜0.05),80 ng/ml( t =2.91,P ＜ 0.05 )had statistical significance.At the concentration of 18.3 ng/ml,triptolide induced PC3 cells apoptosis in a time - dependent manner.Compared with the control group,Anexin-V( + )/PI(-)was(5.42±2.21)％(t =3.52,P ＜0.05)in 6h,(13.51±3.37)％(t =6.53,P ＜0.01) 12h,(29.3 ±4.53)％ ( t =8.74,P ＜0.01) 24 h group separately,and it had statistical significance.RTPCR showed that 18.3 ng/ml triptolide up-regulated the mRNA expression of BAX and PIG3,down-regulated P21 and BCL-2.FAS and CASPASE3 did not show obvious changes.Conclusions Triptolide inhibits the proliferation of PC3 and induces apoptosis,and the changes of BCL-2,BAX,PIG3 and P21 may play an important role in the apoptosis of PC-3 cells.