| Objectives To investigate the inhibition of BMSCs transfected with ADAM17-sh RNA on breast cancer xenografts in nude mouse.Methods Human breast cancer MCF-7 cells were cultured normally.BMSCs were isolated by the whole bone marrow adherence method from 3-week male SD rats,and purified by passage culture.BMSCs were transfected with Lentivirus-mediated(red fluorescent protein expression)ADAM17-sh RNA,and the expression of red fluorescence in BMSCs was observed by laser scanning confocal microscope.Forty female nude mice were subcutaneously implanted MCF-7 cells(0.2ml/mouse,5×107/ml)in the right side of the footpath.After 14 days of implantation,40 nude mice were randomly divided into control group(equal volume PBS),low-dose BMSCs group(1×106/ml BMSCs),highdose BMSCs group(5×106/ml BMSCs),low-dose transfection group(1×106/ml BMSCs transfected with ADAM17-sh RNA)and high-dose transfection group(5×106/ml BMSCs transfected with ADAM17-sh RNA).After 48 h of transfection,BMSCs were digested and made into the cell suspension according to the concentration of each group.Nude mice were injected 0.1ml /mouse by tail vein and repeated injection every 3d for 5 times.Since the 10 th day of implantation,the tumor volume was measured and the general condition of nude mice was observed every 4d.After 30 d of implantation(16d after treatment),all nude mice were sacrificed and tumor sampale were reserved.The expression of ADAM17-m RNA was detected by real-time PCR,and ADAM17 protein was detected by Western blot.Results 1 BMSCs were isolated successfully from SD rats,and BMSCs were purified by passage culture.2 Laser scanning confocal microscope results showed that red fluorescent protein was expressed in most BMSCs,and the transfection rate was up to 80%.3 Tumor nodules were found in all nude mice after 14 d of implantation.4 The implanted tumor volumes of control group,low-dose BMSCs group,high-dose BMSCs group,low-dose transfection group and high-dose transfection group was 786.75±26.20mm3,766.63±29.51mm3,485.06±20.65mm3,362.12±19.70mm3 and 251.56±21.77mm3,respectively.There was significant difference in five groups(F = 806.70,P = 0.00).Compared with control group,the tumor inhibition rate of high-dose BMSCs group,low-dose transfection group and high-dose transfection group was 38.35%,53.97% and 68.02%,respectively.There was no significant inhibition in low-doses BMSCs group.5 Real-time PCR results showed that the expression of ADAM17 m RNA in control group,low-dose BMSCs group,high-dose BMSCs group,low-dose transfection group and high-dose transfection group was 1.00±0.00,0.98±0.02,0.81±0.05,0.31±0.04,0.22±0.03,respectively.Compared with the control group,the expression of ADAM17 m RNA in high-dose BMSCs group,low-dose transfection group and high-dose transfection group was significantly downregulated(P<0.01).6 Western blot results showed that the expression of ADAM17 protein in control group,low-dose BMSCs group,high-dose BMSCs group,low-dose transfection group and high-dose transfection group was 0.71±0.05,0.67±0.02,0.59±0.04,0.45±0.04,0.28±0.06,respectively.The expression of ADAM17 protein in high-dose BMSCs group,low-dose transfection group,high-dose transfection group was significantly lower than that in control group(P<0.01).But there was no statistical change between low-dose BMSCs group and control group in ADAM17 protein(P> 0.05).Conclusions BMSCs transfected with ADAM17-sh RNA and high-dose BMSCs inhibit the growth of breast cancer in nude mice. |
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