Effect Of CMTM6 On The Biological Function Of Colon Cancer Cells

Objectives To investigate the effects of CMTM6 expression on proliferation,migration and invasion ability and EMT of HT29 human colon cancer cells.Methods Lipo-2000 was used to transfer fluorescent si RNA into HT-29 human colon cancer cells.The percentage of cells with fluorescence was observed by inverted fluorescence microscope to evaluate the transfection efficiency.The transfection efficiency was determined to be more than 80%.The experiment was divided into three groups: blank control group,NC-si RNA group and CMTM6-si RNA transfection group(CMTM6-1 group,CMTM6-2 group,CMTM6-3 group).The same amount of PBS was added to the blank control group,Lip-2000 and si RNA of meaningless sequence(NCsi RNA)were added into NC-si RNA group,CMTM6-si RNA transfection group(CMTM6-1 group,CMTM6-2 group,CMTM6-3 group)was added with Lip-2000 to transfect three different synthetic sequences(CMTM6-1,CMTM6-2,CMTM6-3)into HT-29 human colon cancer cells.q RT-PCR was used to detect the interference of CMTM6 m RNA expression and screen the best interference CMTM6-si RNA.Western blot was used to verify the interference of CMTM6 protein expression,and it was confirmed that the expression of CMTM6 was significantly knocked down,and the interference was effective.CCK8 assay was used to detect the changes of cell proliferation after silencing CMTM6 gene expression.Wound scratch assay was used to detect the change of cell migration ability after silencing CMTM6 gene expression.Transwell invasion assay was used to detect the changes of cell invasion ability after silencing CMTM6 gene expression.The expression of EMT-related markers E-cadherin m RNA and N-cadherin m RNA was detected by q RT-PCR method,and the expression of EMT-related markers E-cadherin protein and N-cadherin protein was detected by Western blot method.Results The results of q RT-PCR and Western blot showed that the three transfection sequences had good interference effect on HT-29 human colon cancer cells,and the best interference effect was CMTM6-2 group(P<0.001).The results of CCK8 assay showed that compared with control group and NC-si RNA group,CMTM6-2 group significantly inhibited cell proliferation at 24 h,48h and 72h(P<0.05).The results of wound scratch assay showed that the cell migration ability of CMTM6-2 group was significantly inhibited compared with control group and NC-si RNA group(P<0.05).The results of Transwell invasion assay showed that compared with control group and NC-si RNA group,the invasion ability of CMTM6-2 group was significantly inhibited(P<0.001).The results of q RT-PCR and Western blot showed that the expression of N-cadherin m RNA and protein decreased(P<0.001),while the expression of E-cadherin m RNA and protein increased(P <0.001).Conclusions Silencing CMTM6 can inhibit the proliferation,migration,invasion and EMT of HT-29 human colon cancer cells.Figure19;Table2;Reference 144...