生理学报Acta Physiologica Sinica, April

研究论文

尼氟灭酸对肝癌细胞增殖的影响

田晶1, 陶凌1, 曹云新2, 董玲1, 胡玉珍1, 杨安钢3, 周士胜1,*

1第四军医大学生理学教研室、2免疫学教研室、3生物化学与分子生物学教研室, 西安 710032

摘要: 为了观察氯通道阻断剂尼氟灭酸(NFA)对人肝癌细胞(human hepatoma cell line, HHCC)增殖的影响, 我们将NFA作用于HHCC, 应用细胞计数法及噻唑兰(MTT)比色分析法观察细胞增殖情况; 用流式细胞仪检测细胞周期时相; 并用激光扫描共聚焦显微镜检测[Ca2+]i 的变化。结果发现， NFA使HHCC细胞数及MTT光吸收值(OD)较对照组都显著降低, 去除NFA后, OD值逐渐恢复。经100 μmol/L NFA处理48 h的HHCC细胞G1期细胞比例比对照组明显增高, S期及G2期细胞比例明显低于对照组。细胞外应用NFA (100 μmol/L)使 [Ca2+]i 快速降低, 去除NFA后, [Ca2+]i可恢复。这些结果表明, 尼氟灭酸能抑制细胞增殖, 其机制可能与细胞内信号转导Ca2+/CaM途径被抑制有关。

关键词: 肝癌; 增殖; 氯通道; 钙

中图分类号: Q253; R735.7

Effects of niflumic acid on the proliferation of human hepatoma cells

TIAN Jing1, TAO Ling1, CAO Yun-Xin2, DONG Ling1, HU Yu-Zhen1, YANG An-Gang2 ,ZHOU Shi-Sheng1,*

Departments of 1Physiology, 2Immunology, and 3Biochemistry and Molecular Biology, the Fourth Military Medical University, Xi'an 710032

Abstract: The purpose of this work was to investigate the effects of niflumic acid (NFA), a chloride channel blocker, on the proliferation of human hepatoma cell line (HHCC). Cell proliferation was analyzed by cell count and MTT assay. Cell cycle analysis was carried out by flow cytometry. [Ca2+]i was determined by laser scanning confocal system. It was found that NFA decreased significantly the cell number and the MTT optical density (OD) of HHCC cells, and that the OD value was reversed after washout of NFA. Compared with control, NFA blocked cell cycle progression in G1 phase. Extracellular application of NFA (100 μmol/L) induced a rapid decrease in [Ca2+] i. These findings demonstrate that blockage of chloride channels by NFA induces growth arrest of HHCC in G1 phase, which may be due to the inhibition of Ca2+/CaM-dependent signaling pathways.

Key words: hepatoma; proliferation; chloride channel; Ca2+