Effect of a disintegrin and metalloproteinase 17 short hairpin RNA transfection of bone marrow mesenchymal stem cells on proliferation of human breast cancer MCF-7 cells / 中国综合临床

ABSTRACT

Objective:To investigate the effect of a disintegrin and metalloproteinase 17 (ADAM17) - short hairpin RNA (shRNA) transfection on the proliferation of human breast cancer MCF-7 cells.Methods:Four shRNA sequences (adam17-hsa-297, adam17-hsa-1508, adam17-hsa-1658) targeting ADAM17 were designed, Adam17-hsa-1864) and a negative control (LV10-NC) were used to screen the best inhibition rate of ADAM17 shRNA by qPCR.The experiment was divided into three groups transfection group, meaningless sequence group and control group.RNA was extracted according to routine steps, and then reverse transcripted and amplified.The expression of ADAM17 mRNA was detected by qPCR and the proliferation of MCF-7 cells was measured by MMT method.Results:The results of MTT assay showed that the absorbance values of control group, nonsense sequence group and transfection group were 0.270±0.040, 0.250±0.035 and 0.185±0.080, respectively.There was significant difference between the two groups ( F=3.854, P=0.045). There was no significant difference between the control group and the nonsignificant sequence group ( P>0.05), and the difference between the control group and the transfection group was statistically significant ( P<0.05). There was no significant difference between the nonsignificant sequence group and the transfection group ( P>0.05); the absorbance values of the control group, the meaningless sequence group and the transfection group at 48 h were 0.500±0.057, 0.494±0.086 and 0.311±0.007, respectively, and the differences between the two groups were statistically significant( F=19.42, P<0.001). There were significant differences between transfection group and control group and no significant sequence group (all P<0.05), but there was no significant difference between control group and non significant sequence group ( P>0.05). The absorbance values of control group, nonsense sequence group and transfection group at 72 h were 0.720±0.150, 0.713±0.174 and 0.558±0.071, respectively.There was no Conclusion:BMSC transfected with ADAM17 shRNA could inhibit the proliferation of MCF-7 cells at 24 h and 48 h, while the proliferation of MCF-7 cells decreased at 72 h.