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目的:探讨转染解聚素-金属蛋白酶17-shRNA(a disintegrin and metalloprotease 17-shRNA,ADAM17-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14 d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1×10~6/ml BMMSC)和转染组(注射1×10~6/ml转染ADAM17-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3 d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16 d后处死裸鼠。利用Real-time PCR法检测移植瘤组织ADAM17 mRNA表达,Western blotting法检测移植瘤组织ADAM17蛋白表达。结果:抑瘤实验16 d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)mm~3,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ADAM17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:ADAM17-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。
Objective: To investigate the effect of bone marrow mesenchymal stem cells (BMMSC) transfected with ADAM17-shRNA on nude mice breast cancer MCF-7 cells Inhibitory effect of rat xenografts. Methods: BMMSC of male Sprague-Dawley rats were isolated and cultured by whole bone marrow adherent method. BMMSCs were transfected with lentivirus-mediated ADAM17-shRNA. Thirty nude mice were established MCF-7 breast cancer xenograft model, and the tumor cells were planted successfully 14 days later. The mice were randomly divided into control group (PBS equivalent), BMMSC group (injected with 1 × 10-6 / ml BMMSC) and transfection group (BMMSC injected with 1 × 10-6 / ml ADAM17-shRNA) Each group of 10. On the 15th day of implantation, BMMSCs were injected into the tail vein to inhibit tumor growth (0.1 ml / dose once every 3 days for 5 times). The growth of the nude mice was observed. On the 16th day After the death of nude mice. Real-time PCR was used to detect the expression of ADAM17 mRNA in the xenografts and the expression of ADAM17 in the xenografts was detected by Western blotting. Results: On the 16th day after tumor inhibition, the volume of tumor in control group and BMMSC group was significantly higher than that in transfection group [(787.15 ± 25.95), (767.02 ± 28.98) vs (361.89 ± 19.75) mm ~ 3, all P <0.01] The inhibitory rate of tumor in the BMMSC group was significantly higher than that in the control group (2.57%, 53.89% vs 0.00%, all P <0.05). In control group, the expression level of ADAM17 mRNA in BMMSC group was significantly higher than that in transfected group (1.00 ± 0.01,0.97 ± 0.08 vs 0.30 ± 0.09, all P <0.05); ADAM17 protein expression in control group and BMMSC group was significantly higher than that in transfected group (0.70 ± 0.09,0.68 ± 0.02 vs 0.45 ± 0.05, all P <0.05). CONCLUSION: ADAM17-shRNA can target ADAM17 to transplanted breast cancer in nude mice via BMMSC and exert anti-tumor effect.