The Effects Of ADAM17 Overexpression On The Migration And Invasion Of MCF-7 Breast Cancer Cells

Objectives To survey the influence of ADAM17 gene overexpression on migration and invasion of human MCF-7 breast cancer cells.Methods Human MCF-7 breast cancer cells were routinely cultured,and transfected with.The lentiviral vector carried with red fluorescent protein(RFP)and ADAM17(LV8N-ADAM17).The culture was changed on 24 h after transfection.The expression of RFP in MCF-7 cells was observed by fluorescence inverted microscope on 96 h after transfection.MCF-7 cells were grouped into 3 groups: the transfection group(MCF-7 cells transfected with LV8N-ADAM17),nonsense sequence group(MCF-7 cells transfected with nonsense sequence),and control group(normal MCF-7 cells).After successful transfection,the expression of ADAM17 and EGFR mRNA in different group were detected by real-time PCR.The expression of ADAM17 and EGFR protein in breast cancer cells of each group were detected by western blot.The invasion ability of breast cancer cells in each group was studied by Transwell experiment.The migration capacity of breast cancer cells in each group was detected by cell scratch test.Results 1 The number of multiplicity of infection(MOI)was about 100.On 96 h after transfection,the transfection efficiency was almost 100% observed under the fluorescence inverted microscope.2 Real-time PCR detection showed that the expression levels of ADAM17 mRNA in transfection group,nonsense sequence group and control group were 1.79±0.08,1.16±0.07 and 1.13±0.05,respectively.The expression levels of EGFR mRNA in transfection group,nonsense sequence group and control group were 1.72±0.15,1.21±0.04 and 1.17±0.09,respectively.And the expression level of ADAM17?EGFR mRNA in transfection group were apparently higher than that of control group and nonsense sequence group(P<0.05).3 The results of Western blot showed that the expression of ADAM17 protein were 0.81±0.07,0.52±0.04,0.50±0.02 in the transfection group,nonsense group and control group,respectively.The expression of EGFR protein in the transfection group,nonsense sequence group and control group were 1.43±0.24,0.86±0.06 and 0.88±0.17,respectively.The expression of ADAM17?EGFR protein in transfection group were apparently higher than that of the control group and nonsense sequence group(P<0.05).4 Transwell assay revealed that the number of transmembrane cells in the transfection group,nonsense sequence group,and control group were 101.00±6.19/field,76.35±6.41/field,and 79.20±10.04/field,respectively.The invasion capacity of transfection group was markably higher than the control group and nonsense sequence group,and the difference was statistically significant(P<0.05).5 Cell scratch assay results showed that the rate of scratch healing in transfection group,nonsense sequence group and control group were 70.67%±9.01%,37.00%±2.65% and 39.00%±4.58%,respectively.And the healing rate of scratch in transfection group was obviously higher than that in control group and nonsense sequence group(P<0.05).Conclusions Human MCF-7 breast cancer cells can be transfected with the lentivirus with ADAM17.ADAM17 gene overexpression can enhance the ability of migration and invasion in human MCF-7 breast cancer cells.