细胞内钙在内皮素-1促肺腺癌细胞SPC-A1生长中的作用  

The role of intracellular calcium in endothelin-1-induced proliferation of human lung adenocarcinoma cells SPC-A 1

作　　者：周娟[1] 叶倩君[1] 贾刚[1] 张为民[1] 

机构地区：[1]广州军区广州总医院肿瘤科

出　　处：《肿瘤》2008年第2期108-112,共5页Tumor

基　　金：广东省自然科学基金重点资助项目(编号:04101965);广州市科技攻关资助项目(编号:2005Z3-E0051)

摘　　要：目的：探讨细胞内钙离子（[Ca^2＋]i）在内皮素-1（endothelin-1,ET-1）促肺腺癌细胞SPC-A1生长中的作用及其机制。方法：采用RT-PCR法检测肺腺癌细胞SPC-A1中ET-1及内皮素受体（endothelinreceptor,ETR）的mRNA表达,蛋白印迹法检测SPC-A1细胞中ET-1及ETR的蛋白表达,MTT法检测细胞生长,Fura-2/AM荧光技术检测[Ca^2＋]i。结果：肺腺癌细胞SPC-A1上有ET-1及内皮素受体A（ETAR）及受体B（ETBR）的mRNA及蛋白表达;ET-1（10^-15～10^-8mol/L）具有促进SPC-A1细胞增殖作用,也促进其[Ca^2＋]i浓度升高,作用均呈浓度依赖性;ET-1（10^-10mol/L）促SPC-A1细胞增殖及[Ca^2＋]i浓度升高作用均可被ETAR拮抗剂BQ123（10^-7mol/L）阻断（P〈0.05）,但不受ETBR拮抗剂BQ788（10^-7mol/L）影响（P〉0.05）;去除细胞外Ca^2＋及电压依赖型Ca^2＋通道阻滞剂硝苯地平（1μmol/L）在阻断ET-1升高[Ca^2＋]i浓度作用的同时也抑制ET-1诱导的肺腺癌细胞增殖。结论：ET-1通过ETAR促SPC-A1细胞生长及增加[Ca^2＋]i,细胞外Ca^2＋通过电压依赖型钙离子通道开放内流是ET-1增加[Ca^2＋]i及促肺腺癌细胞SPC-A1增殖的主要机制。Objective：To explore the role of intracellular calcium （[Ca^2＋]i） in endothelin-1 （ET-1）-induced proliferation of human lung adenocarcinoma cells SPC-A 1 and the underlying mechanism. Methods： The mRNA transcription of ET-1 and ET receptor （ETR） were detected by reverse transcription-polymerase chain reaction （RT-PCR） and ET-1 and ETR protein expressions in SPC-A 1 cells were measured by Western blotting. Cell proliferation was measured by （3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide） MTT assay. Intracellular Ca^2＋ concentration was determined by Fura-2/AM fluorescent assay. Results： ET-1 and ETAR and ETBR were expressed in SPC-A 1 cells at mRNA and protein levels. ET-1 （10^-15-10^-8 mol/L） stimulated the proliferation of SPC-A 1 cells in vitro and increased [Ca^2＋]i in a dose-dependent manner （P〈0.05）. The effect of ET-1 （10^-10 mol/L）on the proli-feration and [Ca^2＋]i of SPC-A 1 cells was reversed by a highly selective ET-AR antagonist BQ 123 （10^-7 mol/L, P〈0.05）, but could not be reversed by a highly selective ET-BR antagonist BQ 788（10-7 mol/L, P〉0.05）. Deletion of extracellular Ca^2＋ with edetic acid （EDTA, 0.4 mmol/L） or blockade of voltage-gated calcium channels with nifedipine （1 μmol/L） significantly inhibites ET-1-induced elevation of [Ca^2＋]i and proliferation of SPC-A 1 cells. Conclusion： ET-1 stimulates cell proliferation and increased [Ca^2＋]i by activation of ET-AR in SPC-A 1 cells. Ca^2＋ influx via voltage-gated calcium channel is the main mechanism for [Ca^2＋]i elevation and cell proliferation.

关 键 词：肺肿瘤 内皮缩血管肽1 细胞增殖 细胞 SPC-A1 

分 类 号：R734.2[医药卫生—肿瘤]