Inhibitory Effect Of Na~+-K~+ATP Inhibitor On The Development Of Colorectal Cancer

ObjectiveTo investigate the effect of sodium potassium ATP enzyme(Na~+-K~+ATP)inhibitor on the proliferation,migration and invasion of colorectal cancer cells.And the effect of Na~+-K~+ATP enzyme inhibitor on the effect of Epithelial-Mesenchymaltransition(EMT)ontheepithelialmesenchymal transition(EMT)of cancer cells.It lays a good theoretical foundation for the clinical treatment of colorectal cancer by using Na~+-K~+ATP enzyme inhibitor,which is used in digoxin.MethodsThe MTT experiment was used to determine the IC50 of the colorectal cancer cells in digoxin.Combined use of cell scratch test and Transwell test to determine the prognosis of hecoxin,the proliferation,migration and invasion of cancer cells.The expression level of EMT related markers was detected by protein immunoblotting(Western-blot)in the intervention of digoxin.By using transfection technique(si-RNA)silencing hypoxia inducible factor 1alpha(hypoxia alpha inducible factor-1,HIF-1α),by MTT,Transwell and Western-blot analysis of the effect on the cells,compared with the effect of digoxin on the cells,analysis the mechanism of effects of HIF-1 alpha in colorectal cancer cells and the digoxin the.The prognosis of high symplectic drying and the expression of VEGF after transfection of the virus were detected by q PCR.Result1.MTT experiment for digoxin intervention(in situ HCT116 colon cancer cells)and HT29(non metastatic carcinoma),SW480(orthotopic colon cancer cells(SW620)and lymph node metastasis of colon cancer cells)cell line IC50,and in different concentration and time effect of digoxin intervention,concentration and time correlation between the inhibitory effect of digoxin on node colorectal cancer cell lines,but no significant effect on SW620 cells(P>0.05).2.cell scratch test showed that digoxin could inhibit the migration of HCT116,HT29 and SW620 cells compared with the control group(P<0.05),and had no effect on the growth of NCM460(normal intestinal cells)(P>0.05),but it had no significant inhibitory effect on SW620 cells(P>0.05).3.Transwell invasion test showed that digoxin could inhibit the invasiveness of HCT116,HT29 and SW620 cells(P<0.05),but had no significant inhibitory effect on SW620 cells(P>0.05).4.Western-blot test showed that,compared with the control group HCT116,HT29 and digoxin can inhibit SW480 cells EMT related marker of E-cadherin(E-cadherin)expression(P<0.05),N-cadherin(N-cadherin),SNAIL,Slug and vimentin(vimentin)expression decreased(P<0.05),but the SW620 cells had no significant effect(P>0.05).The effect of 5.virus transfected silent HIF-1-A on the production of cancer cell lines is the same as that produced by digoxin.6.q PCR detection showed that compared with the control group,the expression of VEGF in HCT116,HT29 and SW620 cells decreased after virus transfection and digoxin intervention(P<0.05),but had no significant inhibitory effect on SW620 cells(P>0.05).ConclusionDigoxin can inhibit the proliferation,migration and invasion of HCT116,HT29 and SW480 cells in colorectal cancer,and inhibit the development of EMT,an important biological process of tumor development.The experiment also showed that digoxin had no significant effect on the advanced SW620 cells.Digoxin reduces the expression of VEGF in tumor cells by down regulation of HIF-1-A,thus inhibiting the development of tumor.