·论著·

阻断钙激活钾通道对宫颈癌细胞增殖及钾通道电流的影响

刘玲，聂丹，范凌晔，詹平，钱燕萍，毛熙光

(泸州医学院附属医院，四川泸州646000)

摘要：目的观察钙激活性中电导钾离子通道(IKCa1)被阻断后对宫颈癌HeLa细胞增殖及IKCa1膜电位的影响。方法分别用IKCa1阻断剂克霉唑和RNA干扰法阻断HeLa细胞的IKCa1后，用RT-PCR技术检测HeLa细胞的IKCal mRNA，MTT法检测细胞OD值，膜片钳技术检测细胞IKCa1电流。结果克霉唑阻断IKCa1后，HeLa细胞的IKCa1 mRNA表达降低、IKCa1电流减弱、细胞OD值下降，与对照组相比，P均<0.05。RNA干扰法阻断IKCa1后，Hela细胞的IKCa1 mRNA表达下降、IKCa1电流减弱、细胞OD值降低，与空白对照组和阴性转染组相比，P均<0.05。结论 阻断IKCa1能有效抑制HeLa细胞增殖，下调细胞IKCa1 mRNA的表达，降低其IKCa1电流；IKCa1通过调控HeLa细胞膜电位而影响其信号传递，调控HeLa细胞的增殖。

关键词：离子通道；钾离子通道；钙激活性中电导钾离子通道；钾离子通道阻断剂；克霉唑；小分子干扰RNA技术；宫颈癌细胞；HeLa细胞株

doi：10.3969/j.issn.1002-266X.2015.43.001

中图分类号：R737.33文献标志码：A

基金项目：四川省科技厅应用基础研究基金资助项目(2008JY0014-1)；四川省卫生厅科研基金资助项目(110373)；四川省泸州市科技局科研项目(2013LZLY-J30)。

作者简介：第一刘玲(1981-)，女，硕士，主治医师，主要研究方向为妇科肿瘤、生殖内分泌。E-mail: eye99@163.com

作者简介：通信毛熙光(1965-)，男，硕士，教授，主要研究方向为妇科肿瘤。E-mail: mxg6639@163.com

收稿日期：(2015-07-07)

Effects of blocking intermediate-conductance-Ca2+-activated K+channels

on proliferation of cervical cancer cells and potassium channel current

LIULing,NIEDan,FANLing-ye,ZHANPing,QIANYan-ping,WANGChun-yan,MAOXi-guang

(TheAffiliatedHospitalofLuzhouMedicalCollege,Luzou646000,China)

Abstract:ObjectiveTo investigate the effects of blocking intermediate-conductance-Ca2+-activated K+ channels (IKCa1) on the cell proliferation and the membrane potential changes of IKCa1 in cervical cancer HeLa cells. MethodsIKCal of HeLa cells was blocked with IKCa1 channel inhibitor (clotrimazole, CLT) and RNA interference, respectively. RT-PCR was used to detect the expression of IKCal mRNA in HeLa cells. The OD value of cells was detected by MTT, and IKCal current was detected by the patch clamp technique in HeLa cells. ResultsWhen IKCal was blocked with CLT in HeLa cells, the expression of IKCal mRNA was decreased, IKCal current wakened and the OD value of cells decreased as compared with that of the control group (all P<0.05). When IKCal was blocked by small interfering RNA technology in HeLa cells, the expression of IKCal mRNA reduced, IKCal current wakened, and the OD value of cells decreased as compared with that of the blank group and negative transfection group (all P<0.05). ConclusionsBlocking IKCa1 could inhibit cell proliferation, down-regualte the expression of IKCa1 mRNA and cut down the IKCal current in HeLa cells. IKCa1 could regulate the cell proliferation of HeLa cells by directly regulating the membrane potential and thus affecting its signal transmission.

Key words: ion channel; potassium channel; intermediate-conductance-Ca2+-activated K+channels; potassium channel blocker; clotrimazole; small interfering RNA technology; cervical carcinoma cells; HeLa cell line

目前，宫颈癌的发病机制尚不明确。离子通道是一种成孔蛋白，它通过允许某种特定类型的离子依靠电化学梯度穿过该通道，帮助细胞建立和控制质膜间的微弱电压差，以维持细胞调控信息的正常表达和传递，这些离子通道存在于所有细胞的细胞膜上[1～3]。离子通道包括钾离子通道家族、氯离子通道家族、钙离子通道家族、质子通道家族等。其中钾离子通道的功能异常在肿瘤细胞的生长繁殖过程中发挥了重要作用。……