ADAM17-shRNA转染的骨髓间充质干细胞对裸鼠乳腺癌移植瘤生长的影响

目的:探讨转染解聚素-金属蛋白酶17-shRNA(a disintegrin and metalloprotease 17-shRNA,ADAM 17-shRNA)的骨髓间充质干细胞(bone marrow mesenehymal stem cells,BMMSC)对乳腺癌MCF-7细胞裸鼠移植瘤的抑制效果.方法:全骨髓贴壁法分离并培养3周雄性SD大鼠的BMMSC,利用慢病毒介导的ADAM17-shRNA转染BMMSC.30只裸鼠建立MCF-7乳腺癌移植瘤模型,种植肿瘤细胞14d后建模成功.按照数字表法随机分成对照组(注射等量PBS)、BMMSC组(注射l×106/mlBMMSC)和转染组(注射1×106/ml转染ADAM 17-shRNA的BMMSC),每组10只.在种植细胞第15天开始经尾静脉注射BMMSC进行抑瘤实验(0.1 ml/只,每3d给药1次,共计5次),观察裸小鼠移植瘤的生长情况;抑瘤实验16d后处死裸鼠.利用Real-time PCR法检测移植瘤组织ADAM17 mRNA表达,Westem blotting法检测移植瘤组织ADAM 17蛋白表达.结果:抑瘤实验16d时,对照组、BMMSC组移植瘤体积明显高于转染组[(787.15±25.95)、(767.02±28.98) vs (361.89±19.75)mm3,均P＜0.01];BMMSC组、转染组抑瘤率明显高于对照组(2.57％、53.89％ vs 0.00％,均P＜0.05).对照组、BMMSC组ADAM17 mRNA的表达水平明显高于转染组(1.00±0.01、0.97±0.08 vs 0.30±0.09,均P＜0.05);对照组、BMMSC组ADAM 17蛋白表达水平明显高于转染组(0.70±0.09、0.68±0.02 vs 0.45±0.05,均P＜0.05).结论:ADAM 17-shRNA通过BMMSC介导可将ADAM17靶向归巢至裸鼠乳腺癌移植瘤并发挥抑瘤作用.

Effect of bone marrow mesenchymal stem cells transfected with ADAM17-shRNA on growth of implanted breast cancer in nude mouse

Objective:To investigate the inhibitory effect of bone marrow mesenehymal stem cells (BMMSCs) transfected with ADAM 17-shRNA (adisintegrin and metalloprotease 17-shRNA) on the growth of implanted breast cancer MCF-7 cell xenograft in nude mice.Methods:BMMSCs from 3-week-old male SD rats were isolated and cultured with the whole bone marrow adherence method.BMMSCs were transfected with Lentivirus-mediated ADAM17-shRNA.Breast cancer MCF-7 cell xenograft model was successfully established in 30 nude mice after 14 days implantation of tumor cells.According to the random number table,nude mice were randomly divided into con trol group (equal volume PBS),BMMSCs group (1 × 106/ml BMMSCs) and transfection group (1 × 106/ml BMMSCs transfected with ADAM17-shRNA) with 10 nude mouses in each group.The tumor inhibition test was carried out on the 15th day by injecting BMMSCs into tail vein (0.1 ml/each,administration was carried every 3 days with a to tal of 5 times).The growth of implanted tumor was observed every day.All the nude mice were sacrificed on 16th day after treatment.The expressions of ADAM17 mRNA and ADAM17 protein in rumor tissues were detected by Real-time PCR and Westem blotting,respectively.Results:The volume of implanted tumor in control group,BMMSCs group was significantly larger than that of transfection group ([787.15 ±25.95],[767.02±28.98] vs [361.89±19.75] mm3,all P＜0.01) on D 30.The tumor inhibition rate of BMMSCs group and transfection group was significantly higher than that of control group (2.57％,53.89％ vs 0.00％,all P＜0.05).The expression of ADAM17 mRNA in control group,BMMSCs group was significantly higher than that of transfection group (1.00±0.01,0.97± 0.08 vs 0.30±0.09,P＜0.05).The expression of ADAM17 protein in control group,BMMSCs group was significantly higher than that of transfection group (0.70±0.09,0.68±0.02 vs 0.45±0.05,all P＜0.05).Conclusion:The tropism of ADAM17-shRNA to breast cancer xenograft in nude mice was accomplished by BMMSCs mediation,which may play an anti-tumor effect.