The Effect Of MiR-449a On The Apoptosis And Cell Cycle Of Colorectal Cancer Cell LOVO

Objectives To investigate the expression of miR-449 a in colorectal cancer cells and the effect of miR-449 a on the apoptosis and cell cycle of colorectal cancer cells.Methods Sterile culture of human colorectal cancer cells HCT15,HT29,SW480,LOVO and human colorectal normal mucosal cell line NCM460 in a 37 ?,double gas,100% humidity cell incubator Use qRT-PCR to determine the content of miR-449 a in various cells to pick out the optimal colon cancer cell population;transfect the colon cancer cell line LOVO with liposome Lipofectamine TM 2000 and treat them accordingly: blank group,up-regulated Control group,upregulation group,down-regulation control group,down-regulation group;use qRT-PCR to determine the content of miR-449 a in each group of LOVO cells to verify the effect of transfection;use TUNEL method to measure the apoptosis of the cell kit to determine the withering between different groups Death rate;using cell cycle detection kit,using flow cytometry to determine the inter-cell cycle in different treatment groups;Western blot to determine the changes in apoptosisrelated proteins such as Bcl-2,Bax,and Caspase-3 between LOVO treatments;SPSS Statistic 22.0 data statistical analysis software analyzes the data of each cell experiment group.Results 1 The results of qRT-PCR showed that the expression levels of miR-449 a in human colorectal cancer cells HCT15,HT29,SW480,and LOVO were all lower than those in human colorectal normal mucosal cell line NCM460,and the expression level of LOVO was the lowest(P<0.001).2 The results of qRT-PCR measurement of transfection efficiency showed that in LOVO cells,the expression level of miR-449 a in the up-regulated group was significantly higher than that in the up-regulated NC group,and the expression level in the down-regulated groupwas lower than that in the down-regulated NC group(P<0.05).3 Tunel method detection of apoptosis showed that in LOVO,compared with the NC group and the blank group,the percentage of apoptosis in the up-regulated group was significantly increased(P<0.001).4 Flow cytometry assay: In LOVO,compared with the NC group and the blank group,the percentage of the upregulation group in the G0 / G1 phase increased,which was significantly blocked in the G0 / G1phase(P<0.05).5 Western blot results showed that in LOVO,compared with the NC group and the blank group,the Bcl-2 protein expression level of the upregulation group decreased,while the Bax and Capase-3 expression levels were significantly increased(all P<0.05).Conclusions 1 miR-449 a was up-regulated in colorectal cancer cell lines,and the content of miR-449 a in LOVO was the lowest.2 Up-regulation of miR-449 a can induce apoptosis of colorectal cancer cells and at the same time block cells in G0 / G1 phase.3 miR-449 a may play a role in accelerating the apoptosis of colorectal cancer cells by regulating Bcl-2 protein.Figure 5;Table 8;Reference 170...