靶向抑制PIM-1基因对人前列腺癌裸鼠移植瘤生长的影响  

The effect of PIM-1 silence on the growth of human prostate cancer xenograft tumor in nude mice

作　　者：张晓光[1] 徐勇[2] 张志宏[2] 杨阔[2] 王克明[1] ZHANG Xiao-guang;XU Yong;ZHANG Zhi-hong;YANG Kuo;WANG Ke-ming(Department of Urology, Tianjin Third Central Hospital, Tianjin 300170, China;Tianjin Institute of Urology)

机构地区：[1]天津市第三中心医院泌尿外科,300170 [2]天津市泌尿外科研究所

出　　处：《天津医药》2017年第5期476-480,共5页Tianjin Medical Journal

摘　　要：目的探讨RNA干扰(RNAi)靶向沉默PIM-1基因对人前列腺癌裸鼠移植瘤体内生长的影响。方法12只裸鼠经腋窝下注射前列腺癌细胞(PC-3)建立人前列腺癌裸鼠移植瘤模型。建模后将荷瘤裸鼠随机分为干扰质粒组(瘤体内注射PIM-1特异性RNAi重组质粒P^(PIM1-shRNA-3))、空载体组(空质粒)和阴性对照组(脂质体),每组4只;每隔2 d注射1次,共注射5次。实验期间测量肿瘤体积,绘制肿瘤生长曲线。处理结束后处死动物,测量肿瘤质量,计算抑瘤率。分别采用RT-PCR及Western blot技术检测各组肿瘤标本PIM-1、c-MYC mRNA和蛋白表达情况,免疫组化染色观察各组肿瘤标本PIM-1的表达。结果成功构建人前列腺癌裸鼠移植瘤模型。分组处理后第6天开始,干扰质粒组的肿瘤体积明显小于空载体组和阴性对照组,抑瘤效果明显,同时PIM-1 mRNA及蛋白表达水平均低于空载体组和阴性对照组,PIM-1免疫组化染色亦得出相同结果。干扰质粒组肿瘤组织c-MYC mRNA含量与另2组无明显差异,但c-MYC蛋白的表达明显低于其他2组。结论 RNAi靶向沉默PIM-1可明显抑制前列腺癌移植瘤的生长,并在翻译水平降低c-MYC蛋白的表达,PIM-1可能是前列腺癌基因治疗的有效靶点。To study the effect of PIM-1gene silence by RNA interference(RNAi)on the growth of humanprostate cancer xenograft tumor in nude mice.Methods The xenograft tumor model of human prostate cancer wasestablished by injecting PC-3cells in armpits of12nude mice.After modeling,the nude mice were randomly divided intothree groups:interference plasmid group(injecting with RNAi recombinant plasmid),empty plasmid group and negative control group(liposome every),4mice in each group.Mice were injected every2days for5times.The tumor volumes ofxenografts were measured during experiment,and the curve of tumor growth was drawn accordingly.The quality of tumor wasmeasured,and the inhibitory rate of tumor was calculated at the end of the experiments.The expression levels of PIM-1,c-MYC mRNA and protein in xenograft tumors were detected by real-time PCR and Western blot assay,respectively.Furthermore,immunohistochemistry staining was used to verify the expression of PIM-1.Results The xenograft tumormodel of human prostate cancer was established successfully.The volume of tumor was significantly decreased6days afterthe injection treatment in interference plasmid group than that of empty plasmid group and negative control group.The effectof suppressing tumor growth was remarkable.The expression levels of PIM-1mRNA and protein were down-regulatedsignificantly in interference plasmid group than those of other two groups.The immunohistochemical staining of PIM-1showed the same changes.There was no significant difference in c-MYC protein level between the three groups.Butinterestingly,the c-MYC mRNA level was significantly decreased in interference plasmid group than that of other twogroups.Conclusion The silence of PIM-1gene by RNAi recombinant plasmid can result a significant growth suppressionof the human prostate cancer xenograft tumors in nude mice.The expression of c-MYC gene is down-regulated at translationlevel in the therapeutic group concomitantly.PIM-1may be a promising target of gene therapy for prostate cancer.

关 键 词：前列腺肿瘤 RNA干扰 小鼠 裸 肿瘤异种移植 PIM-1 C-MYC 

分 类 号：R737.25[医药卫生—肿瘤]