Research Of Vacuolization Phenomenon And Related Mechanism Induced By TMEM106A In Hepatocellular Carcinoma Cells

Background and Objective:TMEM106A is affliated to the family of Transmembrane 106, and its specific function is unknown. Existing literature documented the methylaton of the gene in gastric carcinoma cells. It could inhibit the growth of gastric carcinoma cells when its expression was returned, thus it may serve as a tumor supress gene. Besides, it could induce vacuolization in some hepatocellular carcinoma cells. In order to study the specific gene function of TMEM106A, we started from the vacuolization phentype,through retrieval of literature home and abroad, and found the possible relation with pinocytosis. Overexpression of TMEM106A in cells showed the reason of vacuolization,providing information for the futher study.Methods:?. Observe vacuolization phenotype in various hepatocellular carcinoma cell lines like HepG2,HuH-7,PLC/PRF/5.?.Futher study was carried on cell lines showing the vacuolization phenomenon, like the reversibility of vauoles, the start and ending of the cells with vacuoles. Then we used neutral red solution to observe the PH of vauoles, and used electron microscopy to observe the ultrastructure of vacuoles and speculated the possible organelles of the vacuoles. ?. After the possible sources of organelles were found, we searched related signal pathways in literature, and purchased the corresponding inhibitor, finally we used inhibitors experimental to screen the key protein.?. The changes of key proteins were detected by western.Results:?. Vacuolization was observed in HuH-7 and HepG2 cells, but no vauoles was found in PLC/PRF/5 cells. ?. The vacuoles could be stained by neutral red solution,and electron microscopy images observed in vauoles of HepG2 cells showed tabular pseudopodia like structure, which were probably from the membrane surface structure reported to be the key structure of pinosome. ?. The pseudopodia like strcture and growing cell size were found, combined with the lucifer yellow or 70Kd-Rhodamine Dextran found in the vauoles . Then literature was reviewed, and the vacuoles were derived from pinocytosis mostly. Inhibitors like PPP and EHT1864 futher confirmed the relationship with pinocytosis.? . IGF1R signal pathway was detected by western.Conclusion:TMEM106A could induce vacuoles in HepG2 and HuH-7 cells, and this phenomenon could be inhibited by PPP and EHT1864. The vauoles were derived from the endosome system through pinocytosis; TMEM106A may be a novel gene related with pinocytosis.