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钙敏感受体基因多态性E942K通过Ca²⁺和ERK1/2通路促进胃癌细胞增殖

张亚博¹, 杜超², 卢骋³, 董辉³, 吴小翎^1,*

¹重庆医科大学第二附属医院消化科，重庆 400010；²中国人民解放军西部战区总医院消化内科，成都 610038；³陆军军医大学第二附属医院新桥医院消化科，重庆 400037

摘要

本研究旨在探究钙敏感受体(calcium-sensing receptor, CaSR)的单核苷酸多态性(single nucleotide polymorphism, SNP) E942K对胃癌细胞增殖的影响及其机制。对46例胃癌患者和50例正常对照进行基因型检测。应用CaSR E942K突变质粒和野生型质粒转染SGC-7901和HEK-293构建细胞模型，采用CCK8和细胞克隆形成实验检测E942K突变对细胞增殖能力的影响，用钙显影成像技术检测E942K突变对钙信号的影响，并采用Western blot检测E942K突变后下游信号通路关键蛋白ERK1/2和β-catenin磷酸化水平的变化。结果显示，胃癌患者CaSR E942K的突变率显著高于正常对照(P < 0.05)。CaSR E942K突变显著提高了SGC-7901胃癌细胞及HEK-293细胞的增殖能力、细胞内钙信号以及ERK1/2的磷酸化，却不影响β-catenin的磷酸化。以上结果提示，CaSR的E942K突变可能通过增强细胞内钙信号和ERK1/2磷酸化，最终促进了胃癌细胞的增殖，这些结果对胃癌的诊断和靶向治疗具有潜在的临床意义。

关键词： 胃癌; 基因多态性; 细胞增殖; 钙敏感受体; 钙信号; ERK1/2

分类号：R363.2+5

A calcium-sensing receptor polymorphism at E942K promotes the proliferation of gastric cancer cells via Ca²⁺ and ERK1/2 pathways

ZHANG Ya-Bo¹, DU Chao², LU Cheng³, DONG Hui³, WU Xiao-Ling^1,*

¹Department of Digestive System, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China；²Department of Gastroenterology and Hepatology, Chengdu Military General Hospital, Chengdu 610038, China；³Department of Gastroenterology, Xinqiao Hospital, Second Affiliated Hospital of Army Medical University, Chongqing 400037, China

Abstract

The study was designed to investigate the effects and mechanism of a calcium-sensing receptor (CaSR) polymorphism at E942K on the proliferation of gastric cancer cells. Single nucleotide polymorphisms (SNPs) were detected between gastric cancers group and normal controls group by DNA sequence analysis. The cell model was constructed by transfection of E942K mutant plasmid and wild-type (WT) plasmid into SGC-7901 and HEK-293 cells. The effect of E942K mutation on cell proliferation ability was detected by CCK8 and cell clone formation experiments. The effect of E942K mutation on calcium signaling was detected by calcium imaging. Western blot experiments were used to detect changes in phosphorylation levels of key proteins ERK1/2 and β-catenin in downstream signaling pathways after E942K mutation. The results showed that the mutation rate of E942K in gastric cancer group was significantly higher than that in normal control group (P < 0.05). CCK8 and cell clone formation experiments showed that E942K mutation significantly improved the proliferation ability of SGC-7901 gastric cancer cells and HEK-293 cells. E942K mutation enhanced calcium signaling in SGC-7901 and HEK-293 cells. E942K mutation enhanced ERK1/2 phosphorylation without affecting β-catenin phosphorylation. The results suggest that E942K mutation in CaSR may ultimately promote the proliferation of gastric cancer cells by enhancing intracellular calcium signaling and ERK1/2 phosphorylation. These results have potential clinical implications for the diagnosis and targeted therapy of gastric cancer.

Key words: gastric cancer; polymorphism; calcium-sensing receptor; calcium signaling; proliferation; ERK1/2

收稿日期：2019-10-30　　录用日期：2019-05-15

通讯作者：吴小翎　　E-mail: wuxiaoling@cqmu.edu.cn

DOI: 10.13294/j.aps.2020.0038

引用本文：

张亚博, 杜超, 卢骋, 董辉, 吴小翎. 钙敏感受体基因多态性E942K通过Ca²⁺和ERK1/2通路促进胃癌细胞增殖[J]. 生理学报 2020; 72 (3): 274-284.

ZHANG Ya-Bo, DU Chao, LU Cheng, DONG Hui, WU Xiao-Ling. A calcium-sensing receptor polymorphism at E942K promotes the proliferation of gastric cancer cells via Ca²⁺ and ERK1/2 pathways. Acta Physiol Sin 2020; 72 (3): 274-284 (in Chinese with English abstract).