摘要
目的:探讨姜黄素(curcumin)诱导的乳腺癌细胞MCF-7凋亡性途径中氯通道是否参与。方法:MTT检测姜黄素对MCF-7细胞的增殖抑制作用,及用氯通道阻断剂5-nitro-2-(3-phenylpropyl-amino)-benzoate(NPPB)预孵2 h后再加入姜黄素的增殖抑制作用;流式细胞术验证姜黄素诱导细胞的凋亡;采用全细胞膜片钳技术记录加入姜黄素后,细胞氯电流的变化。结果:(1)姜黄素对MCF-7细胞的增殖抑制作用呈现剂量依赖性和时间依赖性。NPPB组与单加入姜黄素组相比,细胞生存率存在明显差异,提示阻滞氯通道可导致姜黄素抑制MCF-7细胞增殖的作用减弱。(2)流式结果表示,加入姜黄素(25μmol·L^-1)24 h后凋亡率33.16%;而姜黄素(25μmol·L^-1)+NPPB(100μmol·L^-1)组凋亡率仅为9.16%,相比对照组(6.26%)、NPPB组(7.31%),氯通道阻断剂明显抑制姜黄素诱导的MCF-7乳腺癌细胞的凋亡,且对早期凋亡的抑制作用明显。(3)全细胞膜片钳技术结果,细胞外灌流姜黄素(25μmol·L^-1)可激活乳腺癌细胞MCF-7氯通道的开放,产生氯电流,翻转电位(-3.18±1.30)mV,外向电流密度(3.16±1.42)pA·pF^-1,内向电流密度为(-2.76±1.38)pA·pF^-1,该电流可被氯通道阻断剂NPPB、DIDS完全抑制,细胞外灌流高渗溶液亦可完全抑制该电流。结论:姜黄素可能通过激活容积敏感性氯通道而诱导细胞的凋亡,氯通道的激活可能在姜黄素诱导肿瘤凋亡通路中起着重要的作用。
OBJECTIVE To investigate the role of chloride channels in curcumin-induced apoptotic pathway in MCF-7 breast cancer cells.METHODS The inhibitory effect of curcumin on the proliferation of MCF-7 cells and the inhibitory effect of curcumin on the proliferation of MCF-7 cells after incubation with 5-nitro-2-(3-phenylpropylamino)-benzoate(NPPB),a chloride channel blocker,was detected by MTT assay.The apoptosis induced by curcumin was verified by flow cytometry.RESULTS(1)Curcumin inhibited the proliferation of MCF-7 cells in a dose-dependent and time-dependent manner.There was a significant difference in cell survival rate between the NPPB group and the curcumin alone group,suggesting that blocking the chloride channel could lead to a decrease in the effect of curcumin on inhibiting the proliferation of MCF-7 cells.(2)Flow results indicated that the apoptotic rate was 33.16%after the addition of curcumin(25μmol·L^-1)for 24 h;while the apoptotic rate was only 9.16%in the curcumin(25μmol·L^-1)+NPPB(100μmol·L^-1)group,compared with the control group(6.26%)and NPPB group(7.31%),the chloride channel blocker significantly inhibited curcumin-induced apoptosis of MCF-7 breast cancer cells,and the inhibition of early apoptosis was significant.The results of whole cell patch clamp technique showed that extracellular perfusion of curcumin(25μmol·L^-1)could activate the opening of MCF-7 chloride channels in breast cancer cells,resulting in a chloride current,with a reversal potential of(-3.18±1.30)mV,an outward current density of(3.16±1.42)pA·pF^-1,and an inward current density of(-2.76±1.38)pA·pF^-1,which was completely inhibited by the chloride channel blockers NPPB and DIDS,and also by the extracellular perfusion of hypertonic solution.CONCLUSION Curcumin induced the cells apoptosis possibly by activating volume-sensitive chloride channels,and the activation of chloride channel may play an important role in curcumin-induced tumor apoptosis.
作者
郭敏
肖苗
叶青
汤莹
GUO Min;XIAO Miao;YE Qin;TANG Ying(Department of Pharmacy,Tongji Hospital,Huazhong University of Science and Technology,Hubei Wuhan 430030,China)
出处
《中国医院药学杂志》
CAS
北大核心
2020年第3期284-288,共5页
Chinese Journal of Hospital Pharmacy
关键词
氯通道
氯通道阻断剂
姜黄素
乳腺癌
MCF-7
chloride channel
chloride channel blocker
curcumin
breast cancer cell
MCF-7
作者简介
郭敏,女,硕士,药师,研究方向:药物新制剂、新剂型及新技术,电话:027-83663231,E-mail:408515048@qq.com;通讯作者:汤莹,女,博士,主管药师,研究方向:中西医结合药理学、药物化学,电话:027-83663084,E-mail:10280921@qq.com
