中国癌症杂志 ›› 2020, Vol. 30 ›› Issue (7): 481-487.doi: 10.19401/j.cnki.1007-3639.2020.07.001

• 论著 • 上一篇    下一篇

ARHGAP4通过调控HK2表达促进肝癌细胞生长的研究

欧阳晓春 1 ,邹叶青 2 ,李玉梅 3 ,丁小兵 3   

  1. 1. 中国人民解放军联勤保障部队第 908 医院神经内科,江西 南昌 330006 ;
    2. 南昌大学第二附属医院江西省重点分子实验室,江西 南昌 330006 ;
    3. 井冈山大学医学部,江西 吉安 343000
  • 出版日期:2020-07-30 发布日期:2020-08-05
  • 通信作者: 丁小兵 E-mail: 563109451@qq.com
  • 基金资助:
    国家自然科学基金(81860530);江西省教育厅科技计划项目(GJJ190564)。

ARHGAP4 promotes growth of hepatocellular carcinoma cells by regulating HK2 expression

OUYANG Xiaochun 1 , ZOU Yeqing 2 , LI Yumei 3 , DING Xiaobing 3   

  1. 1. Department of Neurology, the 908th Hospital of the Chinese People’s Liberation Army Joint Logistic Support Force, Nanchang 330006, Jiangxi Province, China; 2. Key Molecular Laboratory of Jiangxi Province, Second Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China; 3. School of Medicine, Jinggangshan University, Ji’an 343000, Jiangxi Province, China
  • Online:2020-07-30 Published:2020-08-05
  • Contact: DING Xiaobing E-mail: 563109451@qq.com

摘要: 背景与目的:Rho GTP酶活化蛋白4(Rho GTPase activating protein 4,ARHGAP4)与肿瘤的进展密切相关,且对肿瘤细胞恶性增殖具有重要的调节作用。探讨ARHGAP4在肝癌组织中的表达及其对肝癌细胞生长的影响。方法:采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)、蛋白质印迹法(Western blot)及免疫组织化学法分析肝癌组织中ARHGAP4的表达水平,并分析其表达与肝癌临床病理学特征及预后之间的关系。沉默ARHGAP4的表达,采用细胞计数试剂盒(cell counting kit-8,CCK-8)及EdU实验观测肝癌细胞的增殖情况,并检测肝癌细胞中己糖激酶2(hexokinase 2,HK2)的表达水平,进一步分析肝癌组织中HK2的表达及与ARHGAP4的相关性。在稳定低表达ARHGAP4的肝癌细胞中上调HK2的表达,在稳定高表达ARHGAP4的肝癌细胞中沉默HK2的表达,采用Western blot分析HK2的表达情况,采用EdU分析肝癌细胞的增殖能力。结果:肝癌组织中ARHGAP4的表达水平显著高于癌旁组织(P<0.01),且高表达的ARHGAP4与患者肿瘤体积大小及TNM分期密切相关。沉默肝癌细胞中ARHGAP4的表达,肝癌细胞的增殖能力及HK2的表达水平明显减弱(P<0.05);反之过表达ARHGAP4可显著增强HK2的表达及肝癌细胞的增殖能力(P<0.05),且肝癌细胞中ARHGAP4与HK2的表达呈正相关。结论:ARHGAP4通过正向调控HK2的表达进而促进肝癌细胞的生长。

关键词: Rho GTP酶活化蛋白4, 己糖激酶2, 肝癌, 增殖, 凋亡

Abstract: Background and purpose: Rho GTPase activating protein 4 (ARHGAP4) is closely related to tumor progression, and has important regulatory effects on tumor cell malignant proliferation. This study aimed to investigate the expression of ARHGAP4 in hepatocellular carcinoma and its effect on the growth of hepatocellular carcinoma cells. Methods: The expression of ARHGAP4 in hepatocellular carcinoma tissues was analyzed by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR), Western blot and immunohistochemistry. The relationship between the expression of ARHGAP4 and clinicopathological characteristics and prognosis of hepatocellular carcinoma was analyzed. Then, the expression of ARHGAP4 was silenced. Cell counting kit-8 (CCK-8) and EdU experiments were used to observe the proliferation of hepatocellular carcinoma cells, and the expression level of hexokinase 2 (HK2) in hepatocellular carcinoma cells was detected. The expression of HK2 in hepatocellular carcinoma tissues and its correlation with ARHGAP4 were further analyzed. Finally, HK2 expression was up-regulated in hepatocellular carcinoma cells stably expressing low levels of ARHGAP4, while HK2 expression was silenced in hepatocellular carcinoma cells stably expressing high levels of ARHGAP4. Western blot was used to analyze the expression of HK2, and EdU was used to analyze the proliferation of hepatocellular carcinoma cells. Results: The expression levels of ARHGAP4 mRNA and protein in hepatocellular carcinoma tissues were significantly higher compared with adjacent tissues (P<0.01), and the high expression of ARHGAP4 was closely related to the tumor size and TNM stage of patients. Silencing the expression of ARHGAP4 significantly reduced the proliferative ability and HK2 expression level in hepatocellular carcinoma cells (P<0.05). Conversely, overexpression of ARHGAP4 significantly enhanced HK2 expression and cell proliferative ability (P<0.05), and expressions of ARHGAP4 and HK2 in hepatocellular carcinoma were positively correlated. Mechanistically, ARHGAP4 positively regulated the expression of HK2 to affect the growth of hepatocellular carcinoma. Conclusion: ARHGAP4 promotes the growth of hepatocellular carcinoma cells by positively regulating the expression of HK2.

Key words: Rho GTPase activating protein 4, Hexokinase 2, Hepatocellular carcinoma, Proliferation, Apoptosis