| Objective: 1)To investigate the effect of miR-335 on the malignant biological behavior of human intestinal carcinoma HCT-116 cells;2)To preliminarily explore the mechanism of miR-335 affecting the malignant biological behavior of human colorectal cancer HCT-116 cells.Methods:1)Human colorectal cancer HCT-116 cells were cultured in vitro;2)Liposome-mediated transfection of plasmids;3)The subjects were divided into three groups:The Control group(blank group)was the standard human colorectal cancer HCT-116 cells,the Mimics NC group(empty vector group)was the human colorectal cancer HCT-116 cells transfected with no plasmid,and the miR-335 mimics group(transfection group)was the human colorectal cancer HCT-116 cells transfected with miR-335 plasmid;4)The expression of miR-335 in the three groups was detected by reverse transcription quantitative polymerase chain reaction(RT-q PCR)to verify whether the transfection was successful;5)The effects of miR-335 on the proliferation,apoptosis,invasion and migration of human intestinal cancer HCT-116 cells in three groups were detected by MTT assay,flow cytometry,Transwell assay and cell scratch assay,respectively;6)The expression of ZEB2 in human intestinal cancer HCT-116 cells of the three groups was detected by Western blotting;7)The regulation mechanism of miR-335 on ZEB2 was verified by dual luciferase assay in human colorectal cancer HCT-116 cells;8)The software SPSS23.0 was used for statistical analysis of the data.Results: 1)The expression level of miR-335 in the miR-335 mimics group was significantly higher than that in the mimics NC group and Control group,and the difference was statistically significant(P<0.01),there was no significant difference in the expression level of miR-335 between the Mimics NC group and the Control group(P>0.05);2)The OD value of miR-335 mimics group at 24 h,48h and 72 h was lower than that of the mimics NC group and Control group,and the difference was statistically significant at 48 h and 72h(P<0.05),while there was no statistically significant difference in OD values between the Mimics NC group and the Control group at 24 h,48h and 72h(P>0.05);3)The apoptosis rate of miR-335 mimics group was higher than that of the mimics NC group and Control group,and the difference was statistically significant(P<0.01),while there was no significant difference in apoptosis rate between the Mimics NC group and the Control group(P>0.05);4)The number of cell invasion in the miR-335 mimics group was significantly lower than that in the mimics NC group and Control group,and the difference was statistically significant(P<0.01),while there was no significant difference in the number of HCT-116 cells penetrating the membrane between the two groups(P>0.05);5)After 24 h,the width of cell scratches in the miR-335 mimics group was greater than that in the mimics NC group and Control group,and the difference was statistically significant(P<0.01),while there was no statistically significant difference in the scratch width of human intestinal cancer HCT-116 cells in the latter two groups after 24h(P>0.05);6)The expression level of ZEB2 in the miR-335 mimics group was significantly lower than that in the mimics NC group and Control group,and the difference was statistically significant(P<0.01),and there was no significant difference in the expression level of ZEB2 between the Mimics NC group and the Control group(P>0.05);7)The luciferase activity of miR-335+ZEB2 group was lower than that of miR-335+mutant ZEB2 group and other control group,and the difference was statistically significant(P<0.01).Conclusions: 1)miR-335 can inhibit the proliferation,invasion and migration of human intestinal cancer HCT-116 cells,and promote apoptosis;2)The mechanism of miR-335 affecting the malignant biological behavior of human colorectal cancer HCT-116 cells may be related to the binding of miR-335 to downstream target ZEB2 and inhibiting the expression of ZEB2. |
