| Objective: In this study,In this study,a miRNA with regulating HEATR1(HEAT repeat containing 1)gene was predicted and screened,the miRNA was transfected into human gastric cancer cell lines MGC803 and AGS.By observing the cell biological changes between the experimental group(MGC803 and AGS cells transfected with hsa-miR-92a-2-5p mimics)and the control group(MGC803 and AGS cells transfected with negative control mimics),To clarify the effect of hsa-miR-92a-2-5p on proliferation,apoptosis and metastasis of gastric cancer cells by inhibiting HEATR1 gene.Method: Using bioinformatics tools(Target Scan,https://www.targetscan.org/vert＿72)for online statistics filter,choose better combination of five micro RNAs: hsa-miR-92a-2-5p、hsa-miR-20a-2-3p、hsa-miR-183-5p、hsa-miR-506-3p and hsa-miR-556,Five miRNAs mimics were transfected into AGS and MGC803 cells,and the protein expression of target gene was detected by Western blot.The miRNA with the best regulatory effect was selected and transfected into human gastric cancer cell lines MGC803 and AGS.Cell proliferation was observed by CCK-8 proliferation assay,PIFACS cell cycle assay and cell cloning assay,cell apoptosis was detected by flow cytometry,and cell migration and invasion assay were tested.Results:Western Blot experiments were conducted on two kinds of gastric cancer cells(MGC803 and AGS),and the results showed that hsa-miR-92a-2-5p has a good regulatory relationship with HEATR1 gene.After overexpression of hsa-miR-92a-2-5p in AGS and MGC803 cells,colony formation ability of the transfection group was significantly decreased compared with the negative control group(83.000±2.646 vs150.667±7.638,t=14.500,P<0.01),colony formation ability of MGC803 cells in transfection group was significantly decreased compared with negative control group(264.667±9.713 vs 288.333±6.658,t=3.481,P<0.05).The same CCK-8 assay showed that its proliferative ability was also significantly reduced in both types of cells.After96 h of AGS cell culture,the proliferation ability of the transfection group was significantly decreased compared with the negative control group(0.380±0.032 vs0.612±0.019,t=10.621,P<0.01),after 120 h,the proliferation ability of transfection group was significantly decreased compared with negative control group(0.912±0.093 vs 1.590±0.008,t=12.585,P<0.01).After 96 h of culture,the proliferation ability of MGC803 cells in transfection group was significantly decreased compared with that in negative control group(0.588±0.042 vs 0.967±0.025,t=13.691,P<0.01).After 120 h,the proliferation ability of transfection group was significantly decreased compared with negative control group(1.05±2±0.028 vs 1.559±0.013,t=28.550,P<0.01).The apoptosis rate of AGS cells in the transfection group was significantly higher than that in the negative control group(12.667±0.950 vs 5.727±0.482,t=11.279,P<0.01),the apoptosis rate of MGC803 cells in transfection group was significantly higher than that in negative control group(16.133±2.937 vs 5.560±1.361,t=5.658,P<0.01).The subsequent migration and invasion experiments showed that the migration and invasion ability of the transfection group was decreased compared with the negative control group.In AGS cells(360.000±7.937 vs 550.667±10.017,t=25.840,P<0.01;265.333±12.503 vs 455.667±9.292,t=21.163,P<0.01),MGC803 cells(139.667±6.110 vs 241.000±5.568,t=21.232,P<0.01;61.333±9.292 vs 218.333±4.619,t=26.207,P<0.01).Conclusion:This study shows that hsa-miR-92a-2-5p can reduce the expression of HEATR1 gene in gastric cancer cells,inhibit proliferation,invasion and metastasis of gastric cancer cells,and promote apoptosis of gastric cancer cells. |
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