沉默ClC-3氯通道基因对HeLa细胞周期分布的影响

发布时间：2018-03-24 20:31

本文选题：ClC-氯通道　切入点：HeLa细胞　出处：《中国病理生理杂志》2017年02期

【摘要】：目的:探讨沉默HeLa细胞的ClC-3氯通道基因后细胞周期分布的变化及其作用机制。方法:依照siRNA设计原则构建沉默ClC-3基因的ClC-3 siRNA并转染HeLa细胞;实验分为空白对照组(control组)、转染试剂对照组(Lipo组)、阴性对照组(negative siRNA组)和ClC-3 siRNA组。采用real-time PCR检测ClC-3 siRNA的沉默效率;流式细胞术检测细胞周期分布情况;Western blot检测ClC-3蛋白及相关细胞周期蛋白(cyclin)D1、细胞周期蛋白依赖激酶(cyclin-dependent kinase,CDK)4、CDK6、P21和P27等表达。结果:CIC-3 siRNA成功沉默HeLa细胞的ClC-3基因。和其它组相比,ClC-3 siRNA组的细胞周期被阻抑在G_0/G_1期。CIC-3 siRNA组的cyclin D1、CDK4和CDK6蛋白表达水平明显下降,P21和P27蛋白表达水平明显上升。结论:沉默HeLa细胞ClC-3氯通道基因可影响cyclin D1、CDK4、CDK6、P21和27蛋白的表达水平胆抑HeLa细胞周期停滞在G_0/G_1期。
[Abstract]:Objective: to investigate the changes of cell cycle distribution after ClC-3 chloride channel gene silencing in HeLa cells and its mechanism. Methods: ClC-3 siRNA of ClC-3 gene silencing was constructed according to siRNA design principle and transfected into HeLa cells. The experiment was divided into blank control group, transfection reagent control group, negative siRNA group and ClC-3 siRNA group. The silencing efficiency of ClC-3 siRNA was detected by real-time PCR. Flow cytometry was used to detect the distribution of cell cycle. Western blot was used to detect the expression of ClC-3 protein and cyclin D 1, cyclin dependent kinases, cyclin dependent kinase 4, CDK6, P21 and P27. Results the ClC-3 gene of HeLa cells was successfully silenced by siRNA. Compared with the control group, the cell cycle was inhibited in G_0/G_1 phase. CIC-3 siRNA group, the expression level of cyclin D1CDK4 and CDK6 protein decreased significantly. Conclusion: ClC-3 chloride channel gene silencing HeLa cells can affect cyclin D1 CDK4CDK4CDK6P21 and 27 eggs. White expression level inhibited HeLa cell cycle arrest in G_0/G_1 phase.
【作者单位】：广东药科大学基础学院生理学系;暨南大学医学院药理学系;暨南大学医学院生理学系;
【基金】：国家自然科学基金资助项目(No.81272223;No.81273539) 广东药科大学“创新强校工程”-广东高校省级重点平台和重大科研项目:青年创新人才类项目(No.2015cxqx165)
【分类号】：R329.2

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