【摘要】 目的探讨子宫内膜癌组织中钙激活性中电导钾离子通道(IKCal)的表达变化及其在子宫内膜癌细胞增殖中的作用。方法应用蛋白印迹(western blot)法、RT-PCR 技术来研究13份正常子宫内膜及25份子宫内膜癌组织中 IKCal mRNA 和蛋白的表达;利用 IKCal 的阻断剂——克霉唑(clotrimazole),使用小分子干扰 RNA(siRNA)的 RNA 干扰技术和氚胸腺嘧啶核苷掺入试验等观察子宫内膜癌 HEC-1A 细胞增殖的变化。结果子宫内膜癌组织中 IKCal mRNA 的阳性表达率及表达水平(分别为84%、0.89±0.52)明显高于正常子宫内膜组织(分别为8%、0.14±0.12,P 均<0.01)。子宫内膜癌组织中 IKCal 蛋白的阳性表达率及表达水平(分别为80%、1.18±0.41)也明显高于正常子宫内膜组织(分别为15%、0.71±0.26,P 均<0.01)。子宫内膜癌组织中 IKCal mRNA 和蛋白的表达水平与手术病理分期无关(P>0.05),但与病理分级有关(P<0.05)。克霉唑呈剂量和时间依赖性方式阻滞 HEC-1A 细胞的增殖。与转染阴性对照 siRNA 的细胞比较,转染 IKCal siRNA 的 HEC-1A细胞 IKCal 蛋白的表达水平降低,为转染阴性对照 siRNA 的细胞的(48.27±9.07)%。与转染阴性对照 siRNA 的 HEC-1A 细胞比较,转染 IKCal siRNA 能减少细胞的增殖(P<0.05)。结论 IKCal 的异常表达与子宫内膜癌的发展密切相关,降低其表达可减少子宫内膜癌细胞的增殖。更多还原

【Abstract】 Objective To study the expression of intermediate-conductance-Ca²⁺-activated K⁺ （IKCa1）channels in endometrial cancer and its role in regulating proliferation of endometrial cancer cells. Methods Western blot and RT-PCR were used to examine the expression of IKCa1 channels in 13 normal endometrial specimens and 25 endometrial cancer specimens;and RNA interference（RNAi）,[ ³H ] thymidine incorporation,and inhibitor of IKCa1 channel were used to explore the role of IKCa1 channels in regulation of proliferation of endometrial cancer cells HEC-1A.Results The expression rate and level of IKCa1 mRNA in endometrial carcinoma（84%,0.89±0.52）were higher than in normal endometria（8%, 0.14±0.12;P<0.01）.The expression rate and level of IKCa1 protein in endometrial carcinomas（80%, 1.18±0.41）were higher than in normal endometria（15%,0.71±0.26;P<0.01）.Clotrimazole,an inhibitor of IKCal channels known to suppress the function of the channels,caused a both time-and dose- dependent decrease in cell number of HEC-1A cell.Western blot analysis revealed that the IKCa1 level in whole lysates of the cells transfected with target-IKCa1 small interference RNA（siRNA）was（48.27 ±9.07）% of that found in the cells transfected with non-silencing RNA;[ ³H ] thymidine incorporation in HEC-1A cells transfected with target-IKCa1 siRNA was also reduced,siRNA inhibited HEC-1A cell proliferation,compared with the cells transfeeted with non-silencing RNA（P<0.05）.Conclusion The expression of IKCa1 channels may be closely related to the proliferation of endometrial cancer,and down regulation of its expression may suppress its development.更多还原