| Renal Cell Carcinoma(ccRCC)is a common urinary carcinoma,and the most common histological subtype of renal cell carcinoma(RCC)is clear cell renal cell carcinoma(ccRCC),which accounts for 65%-70%among all histological subtypes of RCC.However,molecular mechanisms involving ccRCC pathogenesis and progression are still poorly understood.Currently,HDACs are divided into four classes.HDAC10 is a member of Class Ⅱb family.Our previous study found that HDAC10 was the essential regulator in the clinical prognostic signature based on the HDACs,which could be served as a potential biomarker to estimate the prognosis of ccRCC patients.Moreover,HDAC10 involved in the tumorigenesis and development of various cancers.However,so far,the regulatory mechanism of HDAC10 in mediating the tumorigenesis and development of ccRCC has not been fully elucidated.In this study,we first utilized random forest algorithm to evaluate the impact of the expression of select genes in the constructed signature.Then HDAC10,the highest weight gene,was determined as the target gene.Through bioinformatics prediction,we found that HDAC10 expression was significantly higher in ccRCC samples,which was confirmed by qRT-PCR and immunohistochemistry with clinical samples.We performed MTT,Transwell assay and flow cytometry to test cell proliferation,migration and cell cycle in HDAC10 high expressed-and HDAC10 knockdown ccRCC cells.These results demonstrated that HD AC 10 knockdown significantly inhibited cell proliferation and migration.Cell cycle distribution also showed that inhibition of HDAC 10 induced cell cycle arrest at G1/S phase.These experiments indicated that HDAC 10 played an essential role in promoting ccRCC cell proliferation and migration.Notch signaling pathway plays an essential role in the regulation of cancer cell growth and apoptosis.Previous studies suggested that Notch signaling pathway in cancer cells could promote tumorigenesis driven by tumor suppressor gene phosphatase and tensin homolog(PTEN)loss.However,it is not clear whether aberrant states of Notch pathway exist in ccRCC cells.Still,whether Notch signaling pathway influences the expression of PTEN in ccRCC cells and whether the regulation is associated to the tumorigenesis of ccRCC remain poorly defined.Therefore,Notch signaling pathway and tumor suppressor gene PTEN was used for further investigation.In HDAC 10 knockdown ccRCC cell lines,Western blot analysis was used to detect Notch pathway component notch intracellular domain(NICD),recombination signal binding protein for immunoglobulin kappa J region(RBPJ)and downstream PTEN gene expression at protein levels.These results exhibited that a decrease in NICD and RBPJ in the pathway was observed with HDAC 10 knockdown.Moreover,PTEN was increased in HD AC 10 knockdown ccRCC cells.Then,ccRCC cells were treated with FLI-06,an inhibitor of the Notch signaling pathway,and PTEN was further validated by Western Blot,which revealed that the protein levels of PTEN were remarkably increased.This result indicated that under PTEN engagement,Notch signaling pathway could regulate HDAC10-mediated ccRCC tumorigenesis and development.Considering the deacetylation of HD AC 10,we utilized Quisinostat,an inhibitor of HD AC 10,to treat ccRCC cells,and detected Notch signaling pathway component expression by Western Blot at protein level.We discovered that NICD and RBPJ expression levels were significantly downregulated,while PTEN was upregulated.It is suggested that Notch signaling pathway was regulated through the deacetylation of HD AC 10.In summary,this study identified that HDAC10 could regulate the proliferation,migration and apoptosis of ccRCC.It is clarified that the Notch signaling pathway and PTEN were the main signaling pathway as well as target gene regulating the HDAC10-mediated development of ccRCC.This study further broadens our understanding of the tumorigenesis and development of ccRCC,and will provide new light on targeted therapy for ccRCC. |
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