| Objectives:Although NOTCH1plays a wide-ranging role in controlling cell fate,differentiation and development, its pathological roles in clear cell renal cell carcinoma(CCRCC) are still unclear. In the present study, the expression pattern of NOTCH1wasexamined in CCRCC tissues and the interaction of NOTCH1with the phosphatase andtensin homologue deleted on chromosome10(PTEN)/phosphatidylinositol3-kinase(PI3K)/AKT pathway was investigated in vitro.Methods and Materials:Thirty-six paired CCRCC and adjacent non-neoplastic renal samples wereanalyzed by western blotting and quantitative real-time polymerase chain reaction(qRT-PCR). The alteration of NOTCH1, hairy and enhancer of split1(HES1), PTEN,AKT (phosphorylated at Ser473) in CCRCC cell line (786-O) and the human normalkidney tubule epithelial cell line (HKC) were analyzed by western blotting andqRT-PCR, before and after transfection with siRNA against NOTCH1or the plasmidcontaining the ORF clone of NOTCH1. The effects of NOTCH1signaling pathway oncells proliferation, apoptosis, invasion and migration were detected by MTS assay, flowcytometry analyses and transwell chamber assay respectively.Results:The NOTCH1expression levels were significantly increased in CCRCC tissuescompared with the adjacent non-neoplastic renal samples, while it had no significantassociation with the pathological parameters. NOTCH1signaling cascade wasconstitutively active in human CCRCC cell lines. Blocking NOTCH1signaling resultedin the attenuation of proliferation, invasion and migration, as well as PTENup-regulation with decreased AKT phosphorylation. NOTCH1overexpression had an opposite effect to NOTCH1knockdown.Conclusions:Our findings indicated that NOTCH1receptor expression was up-regulated inCCRCC, and that NOTCH1could regulate PTEN expression and the activity of thePI3K/AKT pathway via HES1in786-O and HKC cell lines. These might provide abasis for the designing new therapeutic strategies for CCRCC.
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