siRNA沉默人宫颈癌基因2抑制胆管癌细胞增殖的实验研究

Experimental study on siRNA silencing human cervical cancer oncogene 2 inhibits proliferation of cholangiocarcinoma cells

目的探讨siRNA抑制人宫颈癌基因2(HCCR-2)表达对胆管癌细胞增殖能力的影响。方法分别采用Pcmv6-AC-GFP/HCCR-2-siRNA慢病毒表达载体及空载体Pcmv6-AC-GFP对照质粒感染人胆管癌RBE细胞,建立siRNA组和对照组。采用Western blot法检测HCCR-2蛋白表达。采用MTT法检测胆管癌细胞体外增殖能力,小鼠皮下移植瘤模型检测胆管癌细胞体内增殖能力。两组实验数据比较采用t检验。结果 siRNA组和对照组人胆管癌RBE细胞的HCCR-2蛋白平均相对表达量分别为0.21±0.03、0.70±0.02。与对照组相比,siRNA组的HCCR-2蛋白相对表达量明显减少(t=-8.06,P<0.05)。感染24 h和48 h后,siRNA组人胆管癌细胞测得的吸光度(A)值分别为0.05±0.01、0.16±0.01,明显小于对照组的0.11±0.02、0.39±0.06(t=-8.80,-11.31;P<0.05)。皮下成瘤模型生长至30 d时,siRNA组小鼠的肿瘤体积为(106±28)mm3,明显小于对照组的(516±24)mm3(t=-29.80,P<0.05)。结论 siRNA沉默HCCR-2基因表达可抑制胆管癌细胞的增殖能力。

Objective To investigate the influence of human cervical cancer gene 2 （HCCR-2） expression inhibited by siRNA on the proliferation of cholangiocarcinoma cells. Methods Pcmv6-AC- GFP/HCCR-2-siRNA lentiviral vector and Pcmv6-AC-GFP control vector were used to infect RBE human cholangiocarcinoma cells to establish the siRNA group and the control group. HCCR-2 protein expression was detected by Western blot. In vitro proliferation of cholangiocarcinoma cells was detected by MTT method and In vivo proliferation of cholangiocarcinoma cells was detected by mice subcutaneous implanted tumor model The experimental data of two groups were compared using t test. Results The average relative expression of HCCR-2 protein in RBE human cholangiocarcinoma cells of the siRNA group and the control group was respectively 0.21±0.03 and 0.70±0.02. Compared with the control group, the relative expression of HCCR-2 protein of the siRNA group was significantly reduced （t=-8.06, P〈0.05）. After 24 h and 48 h infection, the absorbance （.4） measured in human cholangiocarcinoma cells of the siRNA group was respectively 0.05±0.01 and 0.16±0.01, which were significantly lower than 0.11±0.02 and 0.39±0.06 of the control group （t=-8.80, -11.31; P〈0.05）. By the 30th day of subcutaneous implanted tumor grew, the tumor volume of the mice in the siRNA group was （106±28） mm3, which was significantly smaller than （516±24） mm3 of the mice in the control group （t=-29.80, P〈0.05）. Conelusion Expression of HCCR-2 silenced by siRNA may inhibit the proliferation of cholangiocarcinoma cells.