The Effect Of Inhibiting MiR-145 On ADAM17 Expression Of Triple-negative Breast Cancer Cells And Invasion And Proliferation In Vitro

Objective By inhibiting the expression of miR-145,observe the expression of ADAM17 and invasion and proliferation in triple negative breast cancer cells,and explore its mechanism of action.Methods Taking the triple negative breast cancer MDA-MB-231 cell line as the research object,the cells were divided into three groups: control group(normal MDA-MB-231 cells),nonsense sequence group(transfected with miR-145 inhibitor NC MDA-MB-231 cells)and transfection group(MDA-MB-231 cells transfected with miR-145 inhibitor).CCK-8,Transwell,and scratch tests were used to detect the proliferation,invasion,and migration capabilities of the three groups of MDA-MB-231 cells,respectively;real-time quantitative PCR was used to detect the relative expression of miR-145,ADAM17,and EGFR m RNA in the three groups of cells;western blot was used to detect the expression of ADAM17 and EGFR in the three groups of cells.Results Real-time quantitative PCR detection of the relative expression of miR-145 showed that compared with the control group(1.00±0.00)and the nonsense sequence group(0.90±0.09),the relative expression of miR-145(0.64±0.12)in the transfection group was significant.The difference was statistically significant(P<0.05).At 24 h,48h,and 72 h,the transfection group's proliferation ability was significantly higher than that of the control group and the nonsense sequence group,and the difference was statistically significant(P<0.05).The control group and nonsense sequence group at each time point were significantly different.However,there was no statistical difference in cell proliferation between the control group and nonsense sequence group(P>0.05).The results of transwell invasion test showed that the number of perforating cells in the transfection group(127.00±11.45 cells/field),nonsense sequence group(77.00±3.61 cells/field)and control group(74.80±12.11 cells/field),the transfection group the invasion ability of the cells was significantly higher than that of the control group and the nonsense sequence group,and the difference was statistically significant(P<0.05);while the nonsense sequence group was not significantly different from the control group(P>0.05).Scratch test was used to detect the migration ability of breast cancer cells.The results showed that the cell migration ability of the transfection group was significantly stronger than that of the control group and nonsense sequence group,and the difference was statistically significant(P<0.05).Real-time fluorescence quantitative PCR was used to detect the relative expression of ADAM17 and EGFR m RNA in the three groups of breast cancer cells.The expression of ADAM17 m RNA in the transfection group(1.71±0.09)was significantly higher than that in the control group(1.00±0.00)and the nonsense sequence group(1.22±0.40),the difference was statistically significant(P<0.05),while the relative expression of EGFR m RNA was not statistically different between the three groups(P> 0.05).Western blot was used to detect the relative protein content of ADAM17 and EGFR in cells.The relative expression of ADAM17 in three groups of cells(0.327±0.025,0.340±0.096,0.910±0.044),and the relative expression of EGFR protein in the three groups(0.987±0.174,0.887±0.141,1.410±0.277),the protein content of ADAM17 and EGFR in the transfection group were significantly higher than those in the control group and the nonsense sequence group(P<0.05).Conclusion miR-145 inhibitors can act on the ADAM17-EGFR signaling pathway,thereby promoting the proliferation,migration and invasion of triple-negative breast cancer cell MDA-MB-231.Figure 7;Table 3;Reference 208...