天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (11): 1030-1035.doi: 10.11958/20201406

• 细胞与分子生物学 • 上一篇    下一篇

HJURP基因促进肾癌细胞增殖迁移的机制研究

杨柳1，熊吟1，余永红1，胡显锋1，石园园1，江文凛2，李艳兰1，杜艳华1△   

  1. 1武汉市第四医院，华中科技大学同济医学院附属普爱医院老年科（邮编430000），2泌尿外科
  • 收稿日期:2020-05-18 修回日期:2020-08-14 出版日期:2020-11-15 发布日期:2020-11-15

HJURP promotes proliferation and migration of renal cancer

YANG Liu1, XIONG Yin1, YU Yong-hong1, HU Xian-feng1, SHI Yuan-yuan1, JIANG Wen-lin2, LI Yan-lan1, DU Yan-hua1△   

  1. 1 Department of General Surgery, 2 Department of Urology, Wuhan fourth Hospital， Wuhan Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430000, China
  • Received:2020-05-18 Revised:2020-08-14 Published:2020-11-15 Online:2020-11-15

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摘要： 目的　研究HJURP基因对肾癌细胞生物学功能的影响。方法　利用TCGA肾癌数据库分析HJURP基因在肾癌中的表达及对预后的影响，qPCR进一步检测HJURP基因在20例肾癌组织，肾癌细胞株CAKI-1、786O、A498及正常化肾小管细胞HK2中的表达。分别设计HJURP特异性siRNA（si-HJURP组）和对照序列（NC组）转染CAKI-1细胞株，采用细胞克隆形成实验和MTT法检测HJURP基因对肾癌细胞增殖活性影响，流式细胞术检测肾癌细胞周期变化，Transwell实验检测肾癌细胞迁移能力，Western blot检测上皮间质转化（EMT）及AKT通路相关蛋白的表达变化。结果　（1）TCGA数据库中HJURP在肾癌组织中显著高表达，且随着肿瘤分期的升高而增高（P＜0.05）。HJURP高表达组患者总生存率和无病生存率明显较HJURP低表达组患者降低。（2）肾癌组织及肾癌细胞株中HJURP表达均高于癌旁组织及正常肾小管细胞。（3）细胞学实验结果显示，与NC组相比，si-HJURP组CAKI-1细胞增殖活性明显受抑制，G2期细胞比例增高，细胞迁移数量明显下降，EMT相关蛋白N-cadherin表达下降，E-cadherin表达升高，同时AKT通路关键蛋白p-AKT及p-GSK3β明显下调（P＜0.05）。结论　HJURP基因在肾癌中高表达，促进肾癌细胞的增殖及迁移，其可作为肾癌的潜在临床诊治靶点及预后标志物。

关键词: 肾肿瘤；RNA, 小分子干扰；细胞增殖；细胞迁移；细胞运动；细胞周期；HJURP基因

Abstract: Objective　To investigate biological functions of HJURP in renal caner. Methods　TCGA database was used to identify the effect of HJURP on the expression and prognosis in renal cancer. The expression levels of HJURP were detected by real-time polymerase chain reaction (PCR) in 20 samples of renal tumor tissues, renal cell carcinoma cell lines CAKI-1, 786O and A498, and renal tubular cells HK2 . CAKI-1 cell line was transfected with siRNA, and cell colony formation assay and MTT assay were performed to detect the proliferation viability of renal cancer cells in si-HJURP group and control group. Flow cytometry assay was performed to detect the cell cycle changes. Transwell assay were performed to detect cell migration. Western blot assay was used to detect epithelial-mesenchymal transition (EMT) and AKT pathway related proteins. Results　(1)The expression of HJURP was significantly high in renal cancer tissues in the TCGA database, and it increased with the upgrading of tumor stages (P＜0.05). The overall survival rate and disease-free survival rate were significantly decreased in patients with high expression of HJURP than those in patients with low expression of HJURP. (2) The expressions of HJURP were higher in renal cancer tissues and renal cancer cell lines than those in adjacent tissues and renal tubular cells. (3) The results of cytology assay showed that compared with the NC group, the proliferation activity of CAKI-1 cells was significantly inhibited in the si-HJURP group, and the proportion of cells in the G2 phase increased, the number of cell migration decreased significantly, and the expression of EMT-related protein N-cadherin decreased. The expression of E-cadherin increased, and the key proteins of AKT pathway p-AKT and p-GSK3β were significantly down-regulated (P＜0.05). Conclusion　HJURP is highly expressed in renal cancer and promotes the proliferation and migration of renal cancer. It can be used as a potential clinical diagnosis and treatment target and prognostic marker for cancer.

Key words: kidney neoplasms, RNA, small interfering, cell proliferation, cell movement, cell cycle, HJURP gene