| Objective: To investigate the effects of LATS1gene demethylation onthe biological function and Hippo-YAP signaling pathway in human renalcell carcinoma cells.Methods: The expression levels of large tumor suppressor gene1(LATS1) mRNA and downstream cancer gene Yes-associated protein(YAP) mRNA of Hippo-YAP signaling pathway were detected by RT-PCRin human clear cell renal cell carcinoma cell line786-O and humanembryonic kidney cell line HEK-293, the methylation of LATS1in786-Ocells was analyzed by bisulfite sequence-PCR (BSP). After786-O andHEK-293cells were disposed by5-Aza-2’-deoxycytidine (DAC), theexpression levels of mRNA and protein of LATS1and YAP were detectedby RT-PCR and Western Blot respectively in each group, the cell apoptosisand cycle were detected by flow cytometry (FCM) respectively, the cellproliferation inhibition was detected by cell proliferation and toxicitydetection reagent（CCK-8）respectively. Results: Compared with HEK-293, the expression of LATS1mRNAwas significantly decreased (P<0.05), while the expression of YAP mRNAwas markedly increased (P<0.05) in786-O. LATS1was highly methylatedin786-O by BSP; After786-O disposed by DAC, the expression of LATS1mRNA and protein in786-O cells were dramatically increased（P<0.05），but the expression of YAP mRNA and protein was significantly decreased（P<0.05）, the cell cycle was arrested in G1stage (P<0.05), the cellproliferation was obviously inhibited (P<0.05), and the cell apoptosis wasincreased significantly (P<0.05), but changes in HKE-293cells were notsignificant (P>0.05).Conclusion: The demethylation of LATS1gene down-regulates theexpression of YAP gene, inhibits the cell proliferation of786-O and inducesits apoptosis. The methylation of LATS1gene might play an important rolein the occurrences of renal cell carcinoma. |
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