VDR和GLi1在前列腺癌细胞PC-3中的表达及其相关性  

Expression of VDR and GLi1 in prostate cancer cell line PC-3 and its correlation

作　　者：张远东[1] 赵晖[1] 申吉泓[1] 刘孝东[1] 李康健[1] 官润云[1] 

机构地区：[1]昆明医科大学第一附属医院泌尿外科,650032

出　　处：《实用医学杂志》2017年第21期3530-3534,共5页The Journal of Practical Medicine

基　　金：云南省应用基础研究(昆医联合专项基金)(编号:2017FE467-042;2014FB013)

摘　　要：目的初步探讨慢病毒携带sh RNA-VDR载体干扰前列腺癌PC-3细胞中VDR基因对GLi1的影响。方法依照PC-3细胞的培养条件培养细胞;采用荧光定量PCR法和免疫细胞化学SP法检测PC-3细胞中VDR、GLi1两个基因表达情况;对PC-3细胞病毒侵染进行效率评价;运用RT-PCR法检测PC-3细胞VDR基因干扰效果及Gli1转录水平改变情况。结果细胞培养:拍照记录细胞状态:PC-3细胞生长状态良好,以4 d为1周期进行传代;荧光定量PCR法和免疫细胞化学SP法显示VDR、GLi1均在PC-3细胞中表达;慢病毒侵染效率显示为按照1∶10的比例向PC-3细胞加入LV3-NC慢病毒时,细胞侵染效率最好,约为95%左右;RT-PCR显示:VDR-sh RNA慢病毒成功干扰VDR表达,在VDR-sh RNA慢病毒转染72 h后与对照组相比实验组VDR基因转录水平下降85%(P<0.05),同时实验组GLi1基因转录水平与对照组相比上升9%(P<0.05);而当转染96 h后实验组VDR基因转录水平与对照组相比下降99%(沉默),同时实验组GLi1基因转录水平与对照组相比上升248%(P<0.05)。结论所培养的PC-3细胞状态较好;VDR和GLi1基因在PC-3细胞中均表达;慢病毒按照1∶10的比例侵染PC-3效率最高;当VDR被干扰后GLi1表达增高,因而在前列腺癌细胞中维生素D可抑制Hh信号通路可致GLi1表达下调。Objective To investigate the effect of lentivirus carrying sh RNA-VDR vector on GLi1 in pros-tate cancer PC-3 cells. Methods The cells were cultured according to the culture conditions of PC-3 cells.Expression of VDR and GLi1 in PC-3 cells was detected by fluorescence quantitative PCR and immunocytochemistrySP method. The efficiency of PC-3 cell virus infection was evaluated. The effect of VDR gene interference and GLi1 transcription level on PC-3 cells was detected by RT-PCR. Results Cell culture,cell status was recorded and PC-3 cells were in good condition and the passages was 4 days. Fluorescence quantitative and immunocytochemi-cal SP showed that VDR and GLi1 were expressed in PC-3 cells. The virus infection efficiency showed that the in-fection efficiency was about 95% when adding LV3-NC lentivirus to PC-3 cells at 1∶10 ratio. RT-PCR showedthat VDR-sh RNA lentivirus successfully disturbed VDR expression and decreased by 85%(P < 0.05)comparedwith the control group after 72 days of VDR-sh RNA lentivirus transfection. Transcription level of GLi1 gene in theexperimental group increased by 9% compared with the control group(P < 0.05). The transcription level of GLi1 gene in the experimental group increased by 248% compared with the control group(P < 0.05). Conclusion The cultured PC-3 cells were in good condition. VDR and GLi1 genes were expressed in PC-3 cells. Lentivirusshowed highest efficiency in infecting PC-3 at 1∶10 ratio. When VDR was disturbed,the expression of GLi1 in-creased. In prostate cancer cells,vitamin D can inhibit the Hh signaling pathway and cause GLi1 expression downexpression.

关 键 词：PC-3细胞 维生素D受体(VDR) GLI1蛋白 基因干扰 

分 类 号：R737.25[医药卫生—肿瘤]