RNA干扰靶向敲减ObR基因对MCF－7细胞裸鼠移植瘤生长的影响  

Down-regulation of ObR by lentivirus-mediated RNA interference inhibits growth of MCF-7 cells xenograft in a nude mouse model

作　　者：薛荣泉 谷俊朝[2] 杜松涛[2] 俞巍[3] 夏想厚[2] 白志刚[2] 马雪梅[2] 

机构地区：[1]内蒙古自治区人民医院普外科,呼和浩特010017 [2]首都医科大学附属北京友谊医院普外科,北京100050 [3]首都医科大学附属北京天坛医院普外科,北京100050

出　　处：《国际外科学杂志》2012年第4期236-239,共4页International Journal of Surgery

基　　金：北京市自然科学基金（No.7092024）

摘　　要：目的探讨瘤内注射慢病毒介导的瘦素受体的特异性ObR－siRNA对MCF－7人乳腺癌细胞裸鼠移植瘤生长的影响。方法建立荷MCF－7人乳腺癌细胞移植瘤裸鼠模型，共30只，随机分为3组：ObR－siRNA慢病毒干预组、NC－siRNA阴性慢病毒对照组和空白对照组，同时在肿瘤部位周围皮下注射重组人瘦素。隔日测量并记录移植瘤的大小，Real－timePCR和Westernblotting方法检测ObR的mRNA和蛋白水平的表达，测量肿瘤体积，计算肿瘤抑制率。结果成功建立了具有高瘦素微环境的敲减ObR基因的荷MCF－7人乳腺癌细胞移植瘤裸鼠模型。ObR－siRNA慢病毒干预组裸鼠移植瘤的体积与NC．siRNA阴性慢病毒对照组和空白对照组差异有统计学意义（P〈0．01，P〈0．01）。ObR－siRNA慢病毒干预组中的ObR的mRNA和蛋白水平的表达明显降低，相对NC－siRNA阴性慢病毒对照组和空白对照组差异有统计学意义（P〈0．01，P〈0．01），其抑瘤率为88％。结论利用慢病毒载体成功将ObR－siRNA导入移植瘤内，且能显著抑制MCF－7细胞裸鼠移植瘤的生长，提示瘦素受体有可能成为乳腺癌基因治疗的靶点。Objective To investigate the inhibitory effect of lentivirusly-mediated ObR-siRNA on transplanted MCF-7 human breast cancer cells by intratumoral injection. Methods A model of subcutaneous implanted tumor was generated through injecting MCF-7 human breast cancer cells into the nude mice. Thirty established mice with MCF-7 breast cancer cells xenograft were divided into 3 groups randomly, and mice in the experimental group were intratumorally injected with ObR-siRNA lentivirus, while the negative control group and blank control group mice were injected with the same dose of negative lentivirus and normal saline. All mice were subcutaneously injected with recombinant human leptin around the tumor site once a day. Tumor size was blindly measured every other day and the mRNA expression and protein expression levels of ObR in each group were determined. Results Knock- down of ObR-treated xenografted nude mice with a high leptin microenvironment was successfully established. Local injection of ObR-siRNA lentivirus significantly suppressed the established tumor growth in nude mice （P 〈 0. 01, P 〈0.01）. Real time-PCR and Western blotting showed that the mRNA and protein expression of ObR was de- creased in the ObR- siRNA lentivirus group （ P 〈 0. 01, P 〈 0.01 ）. Conclusions Intratumoral injection of recomhi- nant ObR-siRNA lentivirus inhibits the growth of MCF-7 cells xenografts in the nude mice, suggesting that ObR might represent a therapeutic target in the genotherapies of human breast cancer.

关 键 词：乳腺肿瘤 生长抑制物 慢病毒属 瘦素受体 

分 类 号：R737.9[医药卫生—肿瘤]