Effect Of HOXA6 On Cell Proliferation And Apoptosis In Renal Cell Carcinoma

Objective: The aim of this study is to investigate the effect of HOXA6 on cell proliferation and apoptosis in renal cell carcinoma(RCC).Methods: To evaluate the expression level of HOXA6 mRNA and protein in RCC tissues,GSE6344 analysis and immunohistochemical(IHC)were performed.The survival analysis of HOXA6 in RCC was performed with GEPIA.To evaluate the action of HOXA6 on proliferation and apoptosis,RCC cell lines(786-O and 769-P)were transfected with plasmids(Vector,HOXA6,NC and shHOXA6).CCK8,colony formation and EdU were performed to measure cell proliferation,Caspase-Glo and TUNEL were used to detect apoptosis,and flow cytometry(FCM)was used to check cell cycle and apoptosis rate.Bioinformatics analysis was performed to identify the molecular mechanisms of RCC.The Western blotting was performed to measure the protein expression level of PI3K/Akt pathway related proteins.Results: Performed with GSE6344 analysis and IHC,we found that the expression level of HOXA6 mRNA and protein was blocked in RCC tissues.The survival analysis performed with GEPIA revealed that thelower expression of HOXA6 is associated with poor overall survival.As a result,in both 786-O and 769-P cells,compared with the Vector group,the cell proliferation decreased in HOXA6 group;but increased in shHOXA6 group when compared to NC group.As for cell apoptosis,the cell apoptosis rate in HOXA6 group was higher than that in Vector group,while the cell apoptosis rate in shHOXA6 group was lower than that in NC group.Moreover,the FCM results demonstrated that high HOXA6 expression level can block the cell cycle at G0/G1 phase.The result of bioinformatics analysis revealed that DEGs(differentially expressed genes)of RCC were mainly enriched in HIF-1 signaling pathway,focal adhesion,PI3K-Akt signaling pathway and metabolic pathways.The Western blotting showed that in HOXA6 group the expression of PI3 K,p-AKT,MEK,p-ERK and Bcl-2 were suppressed,but the protein expression levels of PTEN,Bax,cleaved-Caspase-3,and cleaved-PARP were increased when compared to Vector group,while the opposite results were observed in shHOXA6 group compared with the NC group.Conclusion: Our study demonstrates that HOXA6 inhibits cell proliferation and promotes apoptosis by inhibiting the PI3K/Akt signaling pathway in RCC.