siRNA沉默人类血红素敏感基因1对膀胱癌T24细胞的影响

目的观察人类血红素敏感基因1（HRG-1）在膀胱正常组织和膀胱肿瘤组织中的表达情况，通过siRNA干扰技术抑制HRG-1在膀胱癌T24细胞的表达水平，观察其生长增殖的变化。方法应用免疫组化SP法检测85例膀胱肿瘤（膀胱乳头状瘤25例，低级别膀胱癌30例，高级别膀胱癌30例）和20例膀胱正常组织石蜡标本中HRG-1表达情况，并进行相关临床病理分析。设计针对HRG-1基因的siRNA，转染膀胱癌124细胞，应用免疫印迹法和RT．PCR分析HRG-1 siRNA-的表达，MTr和流式细胞术（FCM）检测其增殖凋亡的变化。结果正常膀胱组织和膀胱癌组织中HRG-1的表达水平比较差异有统计学意义（P〈0．05）；HRG-1的阳性表达与膀胱肿瘤病例分级、临床分期相关（P〈0．05）；转染siRNA后，HRG-1表达水平下降，MTT和FCM检测发现，细胞增殖曲线受到抑制，HRG-1-siRNA转染组与空载体转染组及未转染组比较，细胞凋亡比例增加，差异均具有统计学意义（P〈0．01）。结论siRNA-HRG-1处理T24细胞后，细胞的增殖受到抑制，同时凋亡比例增加，提示HRG-1基因可能参与了膀胱癌的发病。

Objective To observe the expression change of HRG-1 gene between urinary bladder carcinoma and normal tissues,and to investigate the effect of HRG-1-siRNA on the proliferation and apoptosis of human bladder carcinoma cells.Methods Immunohistochemisty was used to detect the expression of HRG-1 in 85 cases of bladder carcinomas and 20 normal bladder tissues.The siRNA of HRG-1 was designed,synthesized,and transfected into bladder cancer cell line T24.Results The HRG-1 gene expression had significant differences between bladder carcinoma and normal bladder tissues (P ＜ 0.05).The positive expression of HRG-1 gene had significant differences between the pathological grades and clinical stages of bladder carcinomas (P ＜0.05).After treated with siRNA,the expression levels of HRG-1 protein and mRNA in T24 cells decreased obviously (P ＜ 0.05).The apoptosis rate of T24 cells transfected with HRG-1-siRNA was significantly different from control-siRNA group and blank group (P ＜ 0.01).Conclusions The high expression of HRG-1 gene may play an important role in bladder carcinoma,and siRNA targeting HRG-1 can suppress HRG-1 protein expression markedly and enhance apoptosis of T24 cells.