敲低IK1钾通道抑制ClC-3氯通道的表达和功能

发布时间：2018-03-01 11:01

本文关键词： Cl C-氯通道 IK钾通道鼻咽癌膜片钳技术　出处：《中国病理生理杂志》2015年12期 　论文类型：期刊论文

【摘要】：目的:探讨Cl C-3氯通道是否为IK1钾通道的调节靶点,重点研究鼻咽癌细胞IK1钾通道对Cl C-3氯通道功能及蛋白表达的影响。方法:采用siRNA转染技术抑制低分化鼻咽癌上皮细胞(CNE-2Z)IK1基因的表达;real-time PCR技术检测Cl C-3 mRNA的表达;Western blot检测Cl C-3的蛋白表达;细胞免疫荧光结合激光共聚焦显微镜技术检测Cl C-3和IK1蛋白在细胞内分布;全细胞膜片钳记录细胞氯电流。结果:IK1 siRNA可以成功转染CNE-2Z细胞,有效抑制鼻咽癌细胞IK1钾离子通道的表达;用IK1 siRNA抑制鼻咽癌细胞IK1钾离子通道的表达后,Cl C-3的mRNA表达上调而Cl C-3蛋白却表达减少:在低分化鼻咽癌上皮细胞,低渗刺激可激活氯通道,产生一个较大的氯电流,在成功转染IK1 siRNA的细胞,此氯电流明显减弱。结论:敲低IK1钾离子通道可抑制Cl C-3氯离子通道的表达和功能。
[Abstract]:Objective: to investigate whether Cl C-3 chloride channel is the regulatory target of IK1 potassium channel. The effect of IK1 potassium channel on the function and protein expression of Cl C-3 chloride channel in nasopharyngeal carcinoma cells was studied. Methods: siRNA transfection technique was used to inhibit the expression of CNE-2ZOIK1 gene in poorly differentiated nasopharyngeal carcinoma cells. The expression of ClC-3 mRNA was detected by real-time PCR technique. The protein expression of ClC-3 was detected by Western blot. Cellular immunofluorescence and laser confocal microscopy were used to detect the distribution of ClC-3 and IK1 proteins in the cells, and the chloride currents were recorded by whole-cell patch clamp. Results the CNE-2Z cells were successfully transfected with siRNA. After inhibiting the expression of IK1 potassium channel in nasopharyngeal carcinoma cells by IK1 siRNA, the expression of mRNA was up-regulated and the expression of ClC-3 protein was decreased in poorly differentiated nasopharyngeal carcinoma cells. Hypotonic stimulation could activate the chloride channel and produce a large chloride current, which was significantly decreased in the cells successfully transfected with IK1 siRNA. Conclusion: knocking down the potassium channel of IK1 can inhibit the expression and function of Cl C-3 chloride channel.
【作者单位】：暨南大学医学院生理学系;广东药学院基础学院;暨南大学医学院药理学系;
【基金】：国家自然科学基金资助项目(No.81272223)
【分类号】：R739.63

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