Na+/Ca2+交换体抑制剂SN-6和维拉帕米降低肾上腺皮质癌细胞系NCI-H295R醛固酮合成酶表达

doi:10.16352/j.issn.1001-6325.2024.05.0626

基础医学与临床 ›› 2024, Vol. 44 ›› Issue (5): 626-629.doi: 10.16352/j.issn.1001-6325.2024.05.0626

Na⁺/Ca²⁺交换体抑制剂SN-6和维拉帕米降低肾上腺皮质癌细胞系NCI-H295R醛固酮合成酶表达

王宇^1,2, 高寅洁², 任卫东^1,3*, 童安莉^2*

1.河北北方学院研究生院,河北张家口 075000;
2.中国医学科学院北京协和医学院北京协和医院内分泌科国家卫生健康委员会内分泌重点实验室,北京 100730;
3.河北北方学院附属第一医院内分泌科,河北张家口 075000

收稿日期:2024-02-20 修回日期:2024-03-19 出版日期:2024-05-05 发布日期:2024-04-23
通讯作者: ^*tonganli@hotmail.com;15530396532@126.com
基金资助:
国家重点研发计划(2021YFC2501600,2021YFC2501603);国家自然科学基金(82070822);中央高水平医院临床科研业务费(2022-PUMCH-C-028)

Na⁺/Ca²⁺ exchanger inhibitor SN-6 and verapamil reduce aldosterone synthase expression in adrenocortical carcinoma cell line NCI-H295R

WANG Yu^1,2, GAO Yinjie², REN Weidong^1,3*, TONG Anli^2*

1. Graduate School, Hebei North University, Zhangjiakou 075000;
2. Department of Endocrinology, Key Laboratory of Endocrinology of National Health Commission,Peking Union Medical College Hospital, CAMS & PUMC, Beijing 100730;
3. Department of Endocrinology,the First Hospital Affiliated to Hebei North University,Zhangjiakou 075000,China

Received:2024-02-20 Revised:2024-03-19 Online:2024-05-05 Published:2024-04-23
Contact: ^*tonganli@hotmail.com;15530396532@126.com

摘要/Abstract

摘要： 目的研究Na⁺/Ca²⁺交换体(NCX)抑制剂SN-6和钙通道阻滞剂(CCB)维拉帕米对钾离子(K⁺)刺激的肾上腺皮质癌细胞系NCI-H295R(H295R)醛固酮合成酶表达的影响。方法 H295R细胞分为对照组、K⁺(15 mmol/L)处理组、钙通道阻滞剂维拉帕米(verapamil)(10 μmol/L)处理组、SN-6(10 μmol/L)处理组、K⁺+维拉帕米处理组、K⁺+SN-6处理组、维拉帕米+SN-6处理组和K⁺+维拉帕米+SN-6处理组,用实时荧光定量PCR检测醛固酮合成酶(CYP11B2)的mRNA表达,用FLIPR Calcium6检测细胞内钙离子水平。结果与对照组相比,K⁺刺激醛固酮合成酶CYP11B2的mRNA表达(P＜0.001);SN-6和维拉帕米均抑制K⁺刺激的CYP11B2的mRNA表达(P＜0.01); 与K⁺+SN-6组相比,K⁺+SN-6+维拉帕米组更能显著抑制CYP11B2的mRNA表达(P＜0.001)。SN-6和维拉帕米显著降低K⁺刺激的细胞内钙离子水平(P＜0.0001)。结论 SN-6和维拉帕米均抑制K⁺诱导的H295R细胞的醛固酮合成酶的表达;SN-6联合维拉帕米处理,抑制作用更显著。

关键词: SN-6, 维拉帕米, NCI-H295R细胞, 细胞内钙离子, CYP11B2

Abstract: Objective To investigate the effect of Na⁺/Ca²⁺ exchanger inhibitors SN-6 and calcium channel blocker verapamil on potassium-stimulated expression of aldosterone synthase in adrenocortical carcinoma cell line NCI-H295R(H295R). Methods H295R cells were treated in different groups, including control group, K⁺ (15 mmol/L) treated group, verapamil (10 μmol/L) group, SN-6 (10 μmol/L) group, combination of K⁺/verapamil treated group, combination of K⁺/SN-6 treated group, combination of K⁺/verapamil and SN-6 treated group. Real-time PCR was used to detect the mRNA expression of aldosterone synthetase (CYP11B2), and FLIPR Calcium 6 was used to detect the intracellular calcium ion level. Results Compared with the control group, K⁺ stimulated CYP11B2 mRNA expression. Both SN-6 and verapamil decreased K⁺ induced CYP11B2 mRNA expression(P＜0.01). Compared with the K⁺+SN-6 treated group, the K⁺,SN-6 and verapamil treated group all significantly inhibited CYP11B2 mRNA expression (P＜0.001). Both of SN-6 and verapamil significantly reduced the intracellular calcium level stimulated by K⁺ (P＜0.0001). Conclusions Both SN-6 and verapamil inhibit K⁺-induced expression of aldosterone synthase in adrenocortical carcinoma H295R cells. The inhibitory effect is more significant when treated by the combination of verapamil and SN6.

Key words: SN-6, verapamil, NCI-H295R cell, intracellular calcium, CYP11B2

中图分类号:

R586.9

王宇, 高寅洁, 任卫东, 童安莉. Na⁺/Ca²⁺交换体抑制剂SN-6和维拉帕米降低肾上腺皮质癌细胞系NCI-H295R醛固酮合成酶表达[J]. 基础医学与临床, 2024, 44(5): 626-629.

WANG Yu, GAO Yinjie, REN Weidong, TONG Anli. Na⁺/Ca²⁺ exchanger inhibitor SN-6 and verapamil reduce aldosterone synthase expression in adrenocortical carcinoma cell line NCI-H295R[J]. Basic & Clinical Medicine, 2024, 44(5): 626-629.

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Na⁺/Ca²⁺交换体抑制剂SN-6和维拉帕米降低肾上腺皮质癌细胞系NCI-H295R醛固酮合成酶表达

Na⁺/Ca²⁺ exchanger inhibitor SN-6 and verapamil reduce aldosterone synthase expression in adrenocortical carcinoma cell line NCI-H295R

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