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霍利迪连接点识别蛋白调节人肾细胞癌中的细胞凋亡、细胞周期阻滞和活性氧应激。

Holliday junction‑recognition protein modulates apoptosis, cell cycle arrest and reactive oxygen species stress in human renal cell carcinoma.

作者信息

Yuan Jiang-Shui, Chen Zeng-Sheng, Wang Ke, Zhang Zong-Liang

机构信息

Department of Clinical Laboratory, Qingdao Municipal Hospital, Qingdao, Shandong 266011, P.R. China.

Department of Urology, Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.

出版信息

Oncol Rep. 2020 Sep;44(3):1246-1254. doi: 10.3892/or.2020.7656. Epub 2020 Jun 22.

Abstract

Holliday junction recognition protein (HJURP) is involved in the regulation of mortality in various cell types, including renal cell carcinoma (RCC) cells. The specific mechanisms by which HJURP regulates RCC cell apoptosis and the cell cycle have not been previously investigated, to the best of our knowledge. In the present study, the expression of HJURP in RCC tissues and adjacent paracancerous renal tissue, as well as in RCC cell lines, was analyzed using reverse transcription‑quantitative PCR and western blot analysis. The A498 RCC cells were transfected with an HJURP overexpression vector, which resulted in reduced proliferation, as demonstrated using immunofluorescence staining, a Cell Counting Kit‑8 assay and a colony formation assay. Flow cytometry and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labelling assays were used to determine the effect of HJURP on the cell cycle and apoptosis of RCC cells. Proteins associated with the reactive oxygen species (ROS) status were analyzed using western blot analysis. The expression of HJURP was lower in RCC tissues and cells compared with that in the adjacent paracancerous renal tissues and control cells. Furthermore, overexpression of HJURP resulted in a decrease in cell viability and proliferation in vitro. Overexpression of HJURP resulted in cell cycle arrest at the G0/G1 phase, cell apoptosis and an increase in ROS stress. In addition, the phosphorylated/total sirtuin 1 (SIRT1) protein ratio was decreased, whereas the expression of peroxisome proliferator‑activated receptor (PPAR)γ was increased in the HJURP‑overexpressing RCC cells. In clinical practice, decreased HJURP expression may be associated with poor prognosis in patients with RCC. These results suggest that HJURP may regulate cell apoptosis and proliferation in RCC cells and this may be mediated by PPARγ/SIRT1. Thus, HJURP may be used as a predictor of prognosis in patients with RCC.

摘要

霍利迪连接点识别蛋白(HJURP)参与多种细胞类型(包括肾细胞癌(RCC)细胞)的死亡率调节。据我们所知,此前尚未研究过HJURP调节RCC细胞凋亡和细胞周期的具体机制。在本研究中,使用逆转录-定量PCR和蛋白质印迹分析,分析了HJURP在RCC组织、相邻癌旁肾组织以及RCC细胞系中的表达。用HJURP过表达载体转染A498 RCC细胞,免疫荧光染色、细胞计数试剂盒-8检测和集落形成检测结果表明,这导致细胞增殖减少。采用流式细胞术和末端脱氧核苷酸转移酶脱氧尿苷三磷酸缺口末端标记检测,以确定HJURP对RCC细胞的细胞周期和凋亡的影响。使用蛋白质印迹分析与活性氧(ROS)状态相关的蛋白质。与相邻癌旁肾组织和对照细胞相比,RCC组织和细胞中HJURP的表达较低。此外,HJURP的过表达导致体外细胞活力和增殖降低。HJURP的过表达导致细胞周期停滞在G0/G1期、细胞凋亡以及ROS应激增加。此外,在过表达HJURP的RCC细胞中,磷酸化/总沉默调节蛋白1(SIRT1)蛋白比率降低,而过氧化物酶体增殖物激活受体(PPAR)γ的表达增加。在临床实践中,HJURP表达降低可能与RCC患者的预后不良相关。这些结果表明,HJURP可能调节RCC细胞的凋亡和增殖,这可能由PPARγ/SIRT1介导。因此,HJURP可作为RCC患者预后的预测指标。

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