膀胱癌裸鼠移植瘤模型中下调生存素表达的实验研究  

Experimental study of survivin expression in human bladder urothelial carcinoma xenografts in nude mouse model

作　　者：王平[1] 董德平[1] 张建平[1] 朱远源[2] 王桂兰[2] 徐卫东[1] 陈莉[2] 

机构地区：[1]南通大学附属海安医院泌尿外科,江苏省226600 [2]南通大学医学院病理学系

出　　处：《江苏医药》2014年第23期2824-2827,F0002,共5页Jiangsu Medical Journal

基　　金：江苏省产学研联合研究项目(BY2013042-06);南通市社会发展基金资助项目(S11967)

摘　　要：目的探讨靶向生存素(survivin)短发夹样核酸干扰(shRNAi)对裸鼠荷人膀胱尿路上皮癌(BUC)移植瘤的作用。方法构建靶向生存素真核表达质粒(sh-s组)干预人BUC细胞人膀胱尿路上皮癌细胞(T24),同时构建与人类基因序列无同源性的阴性对照表达质粒(sh-T-OFF组)。将构建的质粒分别转染T24细胞。qRT-PCR和Western blot检测细胞内生存素mRNA和蛋白表达。制备荷人BUC裸鼠移植瘤模型;3周后处死裸鼠,免疫组化染色检测瘤组织生存素蛋白的表达和原位末端缺口标记(TUNEL)法检测肿瘤凋亡率(AI)。以丝裂霉素作为对照药物(MMC组)。结果sh-s组移植瘤重量为(2.41±0.03)g,小于sh-T-OFF组的(3.36±0.01)g(P<0.01),大于MMC组的(1.65±0.02)g(P<0.01)。sh-s组和sh-T-OFF组瘤组织中显示明显片状坏死伴炎性细胞浸润。sh-s组裸鼠肿瘤细胞中生存素阳性率为(42.00±2.00)%,低于sh-T-OFF组的(86.00±3.00)%和MMC组的(79.00±2.00)%(P<0.01)。sh-s和MMC组的AI为(33.33±5.77)%和(54.33±4.93)%,高于sh-T-OFF组的(13.38±3.10)%(P<0.05),且MMC组AI高于sh-s组(P<0.05)。结论靶向生存素基因的shRNAi能降低T24细胞和荷人BUC裸鼠移植瘤中生存素表达,抑制肿瘤生长,促进肿瘤细胞凋亡。但其抑癌效果不如MMC。Objective To study the effects of survivin short hair pin sample nucleic acid interference（shRNAi） on the xenografts of bladder urothelial carcinoma（BUC） in nude mice. Methods Eukaryotic expression plasmids for shRNAi expression targeting BUC cell line T24 cells（group sh-s） were constracted. The plasmids with nonhomology human genome sequence were taken as the negative controls（group sh-T-OFF）. The plasmids were transfected into human BUC cell T24 cells. The expressions of survivin protein and mRNA in T24 cells were detected by qRT-PCR and Western blot. The nude mice xenografts models with human BUC were established, which were killed three weeks later for pathological inspection and detecting the expressions of survivin protein in the xenografts by qRT-PCR and TUNEL immunohistochemistry. Mitomycin （MMC） was taken as a positive control drug（group MMC）. Results The xenograft weight of group sh-s was （2.41±0. 03） g,whieh was less than （3.36±0.01） g of group sh-T-OFF（P〈0. 01） ,but more than （1.65±0.02） g of group MMC（P〈0. 01）. There were more necrosis and inflammatory cell reaction in groups of sh-s and sh-T-OFF. The positive rate of survivin in group sh-s was （42.00±2.00）%, which was lower than （86.00±3.00）% in group sh-T-off and （79.00±2.00）% in group MMC（P〈0. 01）. The AI was higher in groups of sh-s and MMC than that in group sh-T-OFF[（33.33±5.77）% and （54. 33± 4. 93） % vs. （l3. 38±3. 10） %3（P〈0.05） ,which was higher in group MMC than that in group sh-s （P%0. 05）. Conclusion The shRNAi targeting survivin gene can significantly reduce the expression of survivin in T24 cells and nude mice BUC xenografts, inhibit the tumor growth and induce the apptosis. But its inhibitory effect on the tumor growth is less than MMC.

关 键 词：膀胱尿路上皮癌 核酸干扰 生存素 丝裂霉素 

分 类 号：R737[医药卫生—肿瘤]