Expression Of Hydrogen Ion Correlative Channels In Hepatic Carcinoma Cells And Its Role In The Growth Of Hepatic Carcinoma Cells

Objective:To investigate the expression of hydrogen ion correlative channels in hepatic carcinoma cells and the role of hydrogen ion correlative channels in the regulation of growth, differentiation and metastasis of hepatic carcinoma cells.Methods:The study was performed in human hepatic carcinoma tissues resected by surgery. Human normal hepatic tissues resected by surgery due to trauma were served as controls. Real-Time RT-PCR and western blot were used to detect the expressions of hydrogen ion correlative channels mRNA and protein in tissues.Human hepatic carcinoma model from subcutaneous to orthotopic transplantion was established in nude mice. The experiments were divided into model group, PPZ (20mg) group, PPZ (40mg) group, PPZ (80mg) group, BM group and DMSO group. Drugs were injected peritoneally once daily. The mice were then killed in one week, two week, three week and four week. The size of tumor was examined, and the histology of tumor tissue was further examined with HE.Results:1. The mRNA expressions of V-ATPase c, V-ATPase a, V-ATPase C and NHE1 in hepatic carcinoma cells were higher than that in normal hepatic cells2. The protein expression of V-ATPase c in hepatic carcinoma cells was obviously higher than that in normal hepatic cells (p<0.01). But, there were no significant differences in protein expression of V-ATPase a, V-ATPase C,V-ATPase B, NHE1 and NHE3 between hepatic carcinoma and normal hepatic cells. There was no protein expression of H+-K+-ATPase alpha, H+-K+-ATPase beta in both hepatic carcinoma and normal hepatic cells.3. The human hepatic carcinoma model from subcutaneous to orthotopic transplantion in nude mice was established successfully in all mice. The volume index of tumor in PPZ (20mg,40mg,80mg) groups were decreased significantly compared with model group after one week,two week, three week and four week (p<0.05).The result indicated that proton pump inhibitor inhibits the growth of hepatic carcinoma cells. Meanwhile, the volume index of BM group and DMSO group showed no difference in one to four week (p>0.05),but both of them were smaller than model group. Histologic examination showed that there is necrosis of large mounts of carcinoma cells in PPZ groups.Conclusion:1. The mRNA and protein expressions of V-ATPase c in hepatic carcinoma cells were enhanced obviously, indicating that V-ATPase c is up-regulated in hepatic carcinoma. This contributes the growth and invasion of hepatic carcinoma cells.2. Proton pump inhibitor pantolazole inhibites the growth of hepatic carcinoma cells and promotes the apoptosis of hepatic carcinoma cells. V-ATPase may be become into new treatment target in hepatic carcinoma pharmacologic treatment.