The Influence Of MiR-214 On The Migration And Apoptosis Of Breast Cancer Cell MCF-7

Objective: To observe the effect of miR-214 on the migration and apoptosis of breast cancer cells MCF-7, to explore its possible mechanism.Methods:1. Detect the expression level of miR-214 in breast cancer cell lines(MDA-MB-231, MCF-7, SK-Br-3) by Real-time PCR, then select the appropriate breast cancer cell lines as the research object of formal experiment.2.miR-214 mimics,miR-214 inhibitors or the independent sequences(NC group) were transfected transiently into breast cancer cells line respectively. The NC group is the negative control and the blank transfection group is the blank control.The transfection efficiency was observed under fluorescence microscope when cultured for 24 h after transfection.3. The miR-214 expression level of breast cancer cell line was investigated by real-time PCR.4. The PTEN protein expression level of breast cancer cell line was by Westem blot.5. The migrate ability of breast cancer cells investigated by Transwell migration assay.6. Detect the apoptosis of breast cancer cells by flow cytometry.Results:1. The miR-214 expression level of the breast cancer cell lines detected by Real-time PCR,which in MCF-7 cells miR-214 expression level(3.268+0.788),lower than that in MDA-MB-231 cells(5.059+0.527),higher than that in SK-Br-3mir-214 cells(1.450+0.390). Compared the three groups in breast cancer cells miR-214 expression levels, the expression level of MCF-7 was moderate and suitable for formal experiment.,the difference was statistical significance(P<0.05).2. After 24 hours transfection, the miR-214 mimics and inhibitors was successfully transfected into human breast cancer MCF-7 cells.The transfection efficiency was over 80%.3.The miR-214 expression level of miR-214 mimics transfection group(7.776+0.891) by Real-time PCR is respectively 2.45 times as much as the control group(316.9+0.981), and the difference was statistical significance(P<0.01).The miR-214 expression level of thecontrol group is 2.57 times as much as the miR-214 inhibitors transfecti-on group(316.9+0.981), the difference was statistical significance(P<0.01).4.Tested by Western blot, the ratio of PTEN/GAPDH protein of miR-214 mimics transfection group(0.142+0.025) is lower than miR-214 inhibitors transfection group(0.741+0.312) and blank control group(0.655+0.468).Compared with the miR-214 mimics, inhibitors transfection group and blank control group, the difference was statis-tically significant(P<0.01).5. Transwell migration assay indicated the MCF-7 cells through the transwell membrane of miR-214 mimics transfection group(216.40+1.053) was higher than miR-214 inhibitors transfection group(26.0+4.47) and blank control group(145.6+10.64). Compared with the miR-214 mimics, inhibitors transfection group and blank control group,the difference was statistically significant(P<0.01).6. The apoptosis rate was 6.84% in the control group by flow cyto-metry. The apoptosis rate of miR-214 mimics transfectiongroup(3.41%) was lower than the control group, the difference was statistically significant(P<0.01). The apoptosis rate of miR-214 inhibitors tran-sfection group(10.28%) was higher than the control group, the diffe-rence was statistically significant(P<0.01).Conclusion:1.miR-214 can promote the migration of human breast cancer MCF-7 cells.2.miR-214 can reduce the apoptosis of human breast cancer MCF-7cells.3.miR-214 may promote the migration of MCF-7 cells by inhibiting the expression of PTEN and decrease the rate of apoptosis.