The Study Of MiR-199b-5p In Breast Cancer

Objective:This study is to explore the clinical significance and correlation through detecting the expression level of miRNA-199b-5p in breast cancer tissue and cells, as well as plasma exosome.it also to investigate the effects of miR-199b-5p on proliferation,migration, apoptosis and invasion in breast cancer cell(HCC1937、MCF-7)through over expression of miR-199b-5p in breast cancer cell(HCC1937、MCF-7), and to explore the relationship between miRNA-199b-5p and DDR1, finally lay the foundation for the clinical treatment and provide a new markers and potential targets for breast cancer.Methods:1, Take miR-199b-5p as the target to synthesize segments of mimic and transfected into HCC1937 and MCF-7 cells by liposome method, while set a the negative control group.2, By the mean of fluorescence and RT-PCR screening of stable transfection cell line.3, With the method of Cell scratch test identify the migration of miR-199b-5p group and negative control group.4,To identify the effect of miR-199b-5p on the migration and invasion in HCC1937 and MCF-7 cells by Transwell assay.5. To detect proliferation ability in HCC1937 and MCF-7 cells with miR-199b-5p mimic transfected compared with negative control by CCK8.6, By the mean of flow cytometry analysis the change of apoptosis rate of HCC1937 and MCF-7 cells with miR-199b-5p mimic transfected compared with negative control.7. Western blotting to detect the expression of DDR1 protein in HCC1937 and MCF-7 cells with transferred with miR-199b-5p mimic and negative control.8. qRT-PCR to detect the expression of miR-199b-5p in breast cancer tissue and cells,as well as plasma exosomes.Results:1, qRT-PCR detection shows that miR-199b-5p mimics can effectively increase the expression of miR-199b-5p in HCC1937 and MCF-7 cells(p<0.05);2,CCK8 assay showed that increase the expression of miR-199b-5p in HCC1937 and MCF-7 cells by mimics resulted in a significant decrease in the proliferation of the HCC1937 and MCF-7 cells(p<0.05).3,Cell scratch test showed increase the expression of miR-199b-5p in HCC1937 by mimics resulted in a significant decrease in the migration rate(p <0.05);4,Flow cytometry showed that the apoptosis rate was increased significantly in miR-199b-5p mimics transfected HCC1937 and MCF-7 cells (p <0.05);5,qRT-PCR results showed that: compared with breast cancer tissues and breast normal tissues. The expression of miR-199b-5p in breast cancer tissues was significantly reduced(p <0.05);6,Westem blotting results showed that: overexpression of miR-199b-5p in HCC1937 and MCF-7 breast cancer cell lines can reduced the expression of DDR1 significantly(p <0.05).Conclusions:1.Over expression of miR-199b-5p can significantly inhibit the proliferation and migration rate, and promote the apoptosis of HCC1937 and MCF-7 breast cancer cells.2.Overexpression of miR-199b-5p in HCC1937 and MCF-7 breast cancer cell lines can significantly reduce the expression of DDR 1.3.Compared with breast normal tissues,the expression of miR-199b-5p in breast cancer tissues was significantly reduced4.Compared with benign breast tumors, the expression of miR-199b-5p in breast cancer peripheral blood exosome is no significant difference.