NOTCH1 functions as an oncogene by regulating the PTEN/PI3K/AKT pathway in clear cell renal cell carcinoma
NOTCH1 functions as an oncogene by regulating the PTEN/PI3K/AKT pathway in clear cell renal cell carcinoma
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NOTCH1 在透明细胞肾细胞癌中通过调节 PTEN/PI3K/AKT 通路发挥癌基因的作用。
DOI：
10.1016/j.urolonc.2011.07.006
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发表时间：
2013-08-01
影响因子：
2.7
通讯作者：
Zhang, Xu
中科院分区：
文献类型：
--
作者：
Liu, Shangwen;Ma, Xin;Zhang, Xu
目的：尽管NOTCH 1在控制细胞命运、分化和发育中发挥广泛作用，但其在肾透明细胞癌（CCRCC）中的病理作用仍不清楚。在本研究中，在CCRCC组织中检测了NOTCH 1的表达模式，并在体外研究了NOTCH 1与10号染色体上缺失的磷酸酶和张力蛋白同源物（PTEN）/磷脂酰肌醇3-激酶（PI3K）/AKT通路的相互作用。通过Western印迹和实时定量聚合酶链反应分析了36对CCRCC和邻近非肿瘤性肾脏样本（qRT-PCR）。通过Western blotting和qRT-PCR分析针对NOTCH 1的siRNA或含有NOTCH ORF克隆的质粒转染前后CCRCC细胞系（786-O）和人正常肾小管上皮细胞系（HKC）中NOTCH 1、多毛和分裂增强子1（HES 1）、PTEN、AKT（在Ser473处磷酸化）的改变。结果：NOTCH 1在CCRCC组织中的表达水平明显高于癌旁组织，但与病理参数无明显相关性。NOTCH 1信号级联在人CCRCC细胞系中是组成性活性的。阻断NOTCH1信号导致增殖、侵袭和迁移的减弱，以及PTEN上调和AKT磷酸化的降低。结论：在786-O和HKC细胞中，NOTCH 1受体表达上调，且NOTCH 1可通过HES 1调节PTEN表达和PI3K/AKT通路活性。为临床设计新的治疗方案提供了依据。(C)2013 Elsevier Inc. All rights reserved.
Objectives: Although NOTCH1 plays a wide-ranging role in controlling cell fate, differentiation, and development, its pathologic roles in clear cell renal cell carcinoma (CCRCC) are still unclear. In the present study, the expression pattern of NOTCH1 was examined in CCRCC tissues, and the interaction of NOTCH1 with the phosphatase and tensin homologue deleted on chromosome 10 (PTEN)/phosphatidylinositol 3-kinase (PI3K)/AKT pathway was investigated in vitro.Materials and methods: Thirty-six paired CCRCC and adjacent non-neoplastic renal samples were analyzed by Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR). The alteration of NOTCH1, hairy and enhancer of split 1 (HES1), PTEN, AKT (phosphorylated at Ser473) in CCRCC cell line (786-O), and the human normal kidney tubule epithelial cell line (HKC) were analyzed by Western blotting and qRT-PCR, before and after transfection with siRNA against NOTCH1 or the plasmid containing the ORF clone of NOTCH. The effects of NOTCH1 signaling pathway on cells proliferation, apoptosis, invasion, and migration were detected by MTS assay, flow cytometry analyses, and transwell chamber assay, respectively.Results: The NOTCH1 expression levels were significantly increased in CCRCC tissues compared with the adjacent non-neoplastic renal samples, while it had no significant association with the pathologic parameters. NOTCH1 signaling cascade was constitutively active in human CCRCC cell lines. Blocking NOTCH1 signaling resulted in the attenuation of proliferation, invasion, and migration, as well as PTEN up-regulation with decreased AKT phosphorylation. NOTCH1 overexpression had an opposite effect to NOTCH1 knockdown.Conclusions: Our findings indicated that NOTCH1 receptor expression was up-regulated in CCRCC, and that NOTCH1 could regulate PTEN expression and the activity of the PI3K/AKT pathway via HES1 in 786-O and HKC cell lines. These might provide a basis for the designing new therapeutic strategies for CCRCC. (C) 2013 Elsevier Inc. All rights reserved.