hERG钾通道介导马钱子碱和士的宁对癌细胞的增殖抑制作用

doi:10.13418/j.issn.1001-165x.2023.6.10

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中国临床解剖学杂志 ›› 2023, Vol. 41 ›› Issue (6) : 684-691. DOI: 10.13418/j.issn.1001-165x.2023.6.10

实验研究

hERG钾通道介导马钱子碱和士的宁对癌细胞的增殖抑制作用

曾纤，周颖，殷雅楠，谢婷，李树基，袁春华*

作者信息 +

hERG potassium channels mediate the anti-proliferative effects of brucine and strychnine on cancer cells

Zeng Xian, Zhou Ying, Yin Yanan, Xie Ting, Li Shuji, Yuan Chunhua*

Author information +

文章历史 +

摘要

目的探讨马钱子碱和士的宁通过阻断hERG通道发挥抗肿瘤作用。方法采用脂质体转染及选择性培养基构建hERG转染CHO细胞和载体转染CHO细胞的稳定细胞株；CCK-8检测共培养24、48、72 h后马钱子碱及其结构类似物对癌细胞增殖的影响，以及共培养48 h后马钱子碱及其结构类似物对hERG-CHO细胞增殖的影响，以载体转染CHO细胞作为对照；流式细胞术检测马HT29、A549细胞周期；免疫印迹法检测HT29、PC9、A549细胞中hERG蛋白表达。结果马钱子碱和士的宁呈时间和浓度依赖性抑制HT29、PC9、A549细胞增殖，这种抗增殖作用与癌细胞中hERG蛋白表达正相关；而马钱子碱和士的宁的氮氧化物仅在高浓度对hERG通道有较弱的阻断活性，对肿瘤细胞也仅在高浓度表现出较弱的增殖抑制作用。马钱子碱对稳定表达hERG的CHO细胞的增殖抑制作用显著高于载体转染的CHO细胞。马钱子碱和士的宁将HT29细胞周期阻滞于G0/G1期，对A549细胞周期没有明显影响。马钱子碱不影响癌细胞的hERG蛋白表达。结论马钱子碱和士的宁对癌细胞的增殖抑制作用可能与hERG通道阻断相关，该研究将为hERG通道阻断剂在肿瘤治疗方面的应用提供重要的指导和依据。

Abstract

Objective To investigate the anti-tumor effect of brucine and strychnine based on their blockage of hERG channels. Methods The vector-transfected and hERG-transfected CHO cells were constructed by Lipofection and selective medium. After treated with brucine and its structural analogues for 24 h, 48 h, and 72 h, the effect in cell proliferation in cancer cells were evaluated by using CCK8 assay. After treated with brucine and its structural analogues for 48 h, the effect in cell proliferation in hERG-transfected CHO cells were evaluated by using CCK8 assay, and the vector-transfected CHO cells were as control. Flow cytometry was used to detect the cell cycle arrest of brucine and strychnine on HT29 and A549 cells. Western blot was used to detect the levels of hERG expression in HT29, PC9, and A549 cells. Results Brucine and strychnine time and dose-dependently inhibited the proliferation of HT29, PC9, and A549 cell lines, and the inhibitory potency was positively correlated with the expression level of hERG protein. Faint anti-proliferation activity was detected at high concentration for brucine N-oxide and strychnine N-oxide which showed weak inhibitory effect on hERG channel and tumor cell only at high concentration. Furthermore, brucine induced more growth suppression on the hERG-transfected CHO cells, when compared with vector-transfected CHO cells. Brucine and strychnine induced G0/G1cell cycle arrest in HT29 cells, and showed no significant effect on the cell cycle of A549 cells. Brucine showed no significant effects on hERG protein expression in the three cell lines. Conclusions Brucine and strychnine displayed their anti-tumor effect may through blocking hERG channels. This study may provide important clues for using the potential pharmacologic effects of hERG inhibitors in cancer treatment.

导出引用

曾纤, 周颖, 殷雅楠, 谢婷, 李树基, 袁春华. hERG钾通道介导马钱子碱和士的宁对癌细胞的增殖抑制作用[J]. 中国临床解剖学杂志. 2023, 41(6): 684-691 https://doi.org/10.13418/j.issn.1001-165x.2023.6.10

Zeng Xian, Zhou Ying, Yin Yanan, Xie Ting, Li Shuji, Yuan Chunhua. hERG potassium channels mediate the anti-proliferative effects of brucine and strychnine on cancer cells[J]. Chinese Journal of Clinical Anatomy. 2023, 41(6): 684-691 https://doi.org/10.13418/j.issn.1001-165x.2023.6.10

中图分类号： R285.5

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