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

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

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 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 AD- AM17- 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 ADAMI7-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 16~ day after treatment. The expressions of ADAM17 mRNA and ADAM17 protein in ttLrnor tissues were detected by Real-time PCR and Western 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 significant- ly 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.