中国癌症杂志 ›› 2016, Vol. 26 ›› Issue (9): 735-742.doi: 10.19401/j.cnki.1007-3639.2016.09.003

• 论著 • 上一篇    下一篇

NCX1在肝癌中的表达、调控Ca2+浓度及其对肝癌细胞增殖和迁移的影响

徐靖宇1,2,江义霞1,谢 睿1,金 海1,文国容1,庹必光1   

  1. 1. 遵义医学院附属医院消化内科,贵州 遵义 563000 ;
    2. 遵义医学院生理学教研室,贵州 遵义 563000
  • 出版日期:2016-09-30 发布日期:2016-10-26
  • 通信作者: 庹必光 E-mail: tuobiguang@aliyun.com
  • 基金资助:
    国家自然科学基金资助项目(81160265);贵州省科技厅、遵义医学院、遵义市科技局科学技术联合基金(黔科合JLKZI2011J46号)。

The expression of NCX1 and its effect on proliferation and migration of hepatocellular carcinoma cells through regulation of intracellular Ca2+

XU Jingyu1,2, JIANG Yixia1, XIE Rui1, JIN Hai1, WEN Guorong1, TUO Biguang1   

  1. 1.Dpeartment of Gastroenterology, Affiliated Hospital, Zunyi Medcial College, Zunyi 563000, Guizhou Province, China; 2.Department of Physiology, Zunyi Medical College, Zunyi 563000, Guizhou Province, China
  • Online:2016-09-30 Published:2016-10-26
  • Contact: TUO Biguang E-mail: tuobiguang@aliyun.com

摘要: 背景与目的:钠钙交换体亚型1(Na+-Ca2+ exchanger isoform 1,NCX1)可通过对细胞Ca2+平衡的调节参与癌症的发生,但是否参与肝癌的发生、发展及其作用机制的研究鲜见报道。该研究旨在探讨NCX1在肝癌中的表达变化,对人肝癌细胞HepG2增殖、迁移能力的影响及其可能的机制。方法:运用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)和蛋白[质]印迹法(Western blot)检测NCX1 mRNA及蛋白在人正常肝细胞株LO2、肝癌细胞株HepG2、人正常肝组织和原发性肝细胞癌患者癌组织中的表达。采用共聚焦显微镜技术观察NCX1在细胞外无钠溶液刺激活化后,对正常肝细胞LO2及肝癌细胞HepG2中钙离子浓度的调控。采用MTT法、细胞划痕实验检测NCXl特异性的阻断剂KB-R7943对人肝癌细胞HepG2增殖、迁移的影响。结果:在肝癌细胞株HepG2和肝细胞癌组织中,NCX1 mRNA和蛋白质的表达均高于正常肝细胞株LO2和正常肝组织(P<0.05)。共聚焦显微镜实验发现,细胞外无钠溶液可以激活细胞内钙升高,正常肝细胞LO2及肝癌细胞HepG2细胞内钙离子浓度均增高,但肝癌细胞HepG2细胞内钙离子增高的幅值明显高于LO2细胞(P<0.05),NCXl特异性阻断剂KB-R7943可以显著阻断胞外无钠诱导的细胞内钙离子升高(P<0.05)。KB-R7943可以显著抑制肝癌细胞HepG2的增殖及迁移(P<0.05)。结论:原发性肝癌中NCX1的表达量上调,NCX1的活化可以调节细胞内钙变化,抑制NCX1的活性可以进一步抑制肝癌细胞的增殖和迁移。这提示NCX1可能在原发性肝癌的发生、发展中起了重要的作用。

关键词: 钠钙交换体亚型1, 钙离子, 肝细胞癌, 细胞增殖, 细胞迁移, KB-R7943

Abstract: Background and purpose: Previous studies have suggested Na+-Ca2+ exchanger isoform 1 (NCX1) as a key component of calcium homeostasis was involved in the tumorigenesis. However, the role of NCX1 and calcium signal in tumorigenesis of hepatocellular carcinoma (HCC) has not been explored. This study aimed to investigate the effect of NCX1 on cell proliferation and migration of HCC HepG2 cells in vitro and the possible mechanism. Methods: Both the real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot were applied to assess the expression of NCX1 mRNA and protein in normal hepatic cells (LO2), HCC cell line (HepG2), human normal hepatic tissues and hepatocellular carcinoma tissues. The change of intracellular calcium signal in LO2 and HepG2 cells via activated NCX1 channel in the presence or absence of Na+ was examined by a confocal laser scanning microscope. The effects of NCX1 special inhibitor KB-R7943 on cell proliferation and migration of HepG2 cells were measured by MTT and cell scratch test. Results: Both mRNA and protein expression of NCX1 were higher in HCC tissues and cell line HepG2 than in the normal tissues and cell line LO2 (P<0.05). The activation of NCX1 channel induced a slight rise in cytoplasmic Ca2+ concentration ([Ca2+]cyt) in normal cells, but caused a marked increase in cancer cells. And the NCX1 activation induced intracellular calcium increase was significantly reversed by NCX1 inhibitor KB-R7943 (P<0.05). Both NCX1-mediated proliferation and migration of HepG2 were also significantly attenuated by the KB-R7943 (P<0.05). Conclusion: NCX1 is up-regulated in HCC cells and tissues. The activation of NCX1 mediates intracellular calcium homeostasis. The inhibition of NCX1 activity can suppress the proliferation and migration of HepG2 cells. It is suggested that NCX1 may be involved in the development and progression of HCC.

Key words: Na+-Ca2+ exchanger isoform 1, Calcium, Hepatocellular carcinoma, Proliferation, Migration, KBR7943