siRNA下调高迁移率族蛋白 A2表达对前列腺癌细胞增殖凋亡的影响

摘要

Objective To study the effect of high mobility group A2(HMGA2) down‐regulation by small interference RNA(siRNA) on the proliferation and apoptosis of the prostate cancer cell line DU145 .Methods With LipofectamineTM 2000 ,HMGA2‐siRNA(group A) and negative control‐siRNA (group B) were transfected into DU145 cells and then the expressions of HMGA2 mRNA and protein were detected by RT‐PCR and Western blot ,respectively .The proliferation and apoptosis of DU145 cells were detected by cell counting kit‐8(CCK‐8) and flow cytometry ,respectively .The results were compared to those of blank controls(group C) .Results The expression of HMGA2 mRNA in group A was 0.562 ± 0.067 ,which was lower than 0.858 ± 0.043 in group B and 0.876 ± 0.041 in group C . So did the cell proliferation(P<0 .05) .The apoptosis rate of DU145 cells in group A was (14.180 ± 0.395)% ,which was higher than (7.513 ± 0.476)% in group B and (8.267 ± 0.379)% in group C (P<0 .05) .Conclusion Silencing HMGA2 gene can effectively inhibit the expressions of HMGA2 mRNA and protein ,decrease the ability of proliferation ,and promote the apoptosis of DU145 cells .%目的：观察小干扰RNA（siRNA）靶向下调高迁移率族蛋白A2（HMGA2）基因表达对人前列腺癌DU145细胞增殖和凋亡能力的影响。方法采用脂质体LipofectamineTM 2000将HMGA2‐siRNA（siRNA组）和阴性对照siRNA（阴性对照组）分别转染至DU145细胞中；空白组无特殊处理。采用RT‐PCR及Western blot 法分别检测 HMGA2 mRNA 和蛋白的表达；细胞增殖‐毒性检测（CCK‐8）法检测DU145细胞的增殖；流式细胞术检测细胞凋亡。结果 siRNA组 HMGA2 mRNA的相对表达量为0.562±0.067，低于阴性对照组的0.858±0.043和空白组的0.876±0.041（P＜0．05）。siRNA组细胞的增殖能力也低于其他两组（P＜0．05）。转染后48 h ，siRNA组DU145细胞的凋亡率为（14.180±0.395）％，高于阴性对照组的（8.267±0.379）％和空白组的（7.513±0.476）％（P＜0．05）。结论 HMGA2基因特异性siRNA可抑制DU145细胞增殖，促进细胞凋亡。