The Research Of ARK5Knocked Downand Its Biological Role On SMMC-7721Cells

OBJECTIVE: Firstly to detect the different expression of ARK5inhepatocellular cacinoma tissues and cells, and then to investigate thebiological effect of hepatoma cells.METHODS: ARK5mRNA and protein were determined by RT-PCRand Western blot between30pairs of hepatocellular carcinoma (HCC)tissues and paracarcinoma tissues, then between hepatoma SMMC-7721cells and hepatic cells (LO2). Small interfering RNA (siRNA) of ARK5and Negative control (NC) siRNA were constructed and transfected intoSMMC-7721cells. Interference group, negative group and blank groupwere set up. MTT assay was used to investigated the proliferation oftransfected cells, Flow cytometry (FCM) was used to examine apoptosis intransfected cells. Transwell assay was used to determined the effect oftransfected cells on migration and invasion.RESULTS: PCR and Western blot showed that ARK5mRNA andprotein in HCC tissues were both significantly higher than those inparacarcinoma tissues and the expression of ARK5in SMMC-7721cellswas also higher than which in LO2cells. MTT assay demonstrated that the cell growth inhibition rates (%) of transfected cells in interference group at24,48and72h were19.39±5.42、23.19±0.53and20.74±1.23, there weresignificant differences compared with negative and blank group (P＜0.01).The results of FCM indicated that the apoptotic rate of the cells ininterference group was15.017±0.945, there were significant differencescompared with negative and blank group (P＜0.01). Transwell assayshowed the number of metastatic cells was139±21in interference group,which in negative group and blank group were209±16and203±15respectively. The number of invasive cells was35±4in interference group,which in negative group and blank group were58±9and61±7respectively.There were statistically significant differences﹙P＜0.05﹚.CONCLUSION: ARK5expression significantly increased in HCC,inhibited the expression of ARK5could effectively suppress hepatoma cellgrowth, induce apoptosis and inhibit migration and invasion, So ARK5maybe act as a cancer-promoting gene and induce hepatocellularcarcinogenesis.