Synergistic inhibitory effect of resveratrol and TK/GCV therapy on melanoma cells
Synergistic inhibitory effect of resveratrol and TK/GCV therapy on melanoma cells
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白藜芦醇与TK/GCV治疗对黑色素瘤细胞的协同抑制作用
DOI：
10.1007/s00432-020-03203-z
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发表时间：
2020-04-03
影响因子：
3.6
通讯作者：
Tan, Yuhui
中科院分区：
文献类型：
--
作者：
Chen, Yuan;Li, Hong;Tan, Yuhui
目的探讨白藜芦醇对TK/GCV自杀基因系统旁观者效应的协同作用。方法采用MTT法和高容量筛选法检测白藜芦醇对B16细胞生长的影响以及白藜芦醇与GCV合用时的协同作用。采用流式细胞术结合荧光示踪技术和荧光显微镜检测白藜芦醇对GJIC功能的影响，采用Western blotting检测间隙连接蛋白的表达。使用移植的黑素瘤模型在体内测试白藜芦醇与TK/GCV的协同杀伤作用。结果白藜芦醇可增强B16细胞GJ功能，上调Cx 32和Cx43蛋白表达。白藜芦醇与GCV协同杀伤混合的B16黑色素瘤细胞（20% TK+细胞和80% TK-细胞），提高TK-细胞的凋亡率（TK系统的旁观者效应），这种协同作用可被GJ抑制剂阿加逆转。在体内，当B16细胞与30%TK + B16细胞混合时，观察到与GCV或白藜芦醇单独处理相比，白藜芦醇联合GCV后肿瘤重量和体积显著减小。结论白藜芦醇可协同增强TK/GCV自杀基因系统对黑色素瘤B16细胞和移植性黑色素瘤的杀伤作用。它可能是TK/GCV治疗的一种有前途的辅助药物。
Purpose To investigate the synergistic effect of resveratrol on the bystander effect of TK/GCV suicide gene system in melanoma cells. Methods The effect of resveratrol on the growth of B16 cells and the synergistic effect of resveratrol with or without GCV were detected by MTT assay and high content screening assay. The effect of resveratrol on GJIC function was detected by flow cytometry combined with fluorescence tracer and fluorescence microscope, and the expression of gap junction protein was detected by western blotting. Synergistic killing effect of resveratrol plus TK/GCV was tested in vivo using transplanted melanoma model. Results In vitro, resveratrol can enhanced GJ function and upregulated Cx32 and Cx43 protein expression in B16 cells. Resveratrol synergized with GCV to kill mixed B16 melanoma cells (20% TK+ cells and 80% TK- cells) and to improve apoptosis rate of TK- cells (the bystander effect of TK system), and the synergistic action was reversed by the GJ inhibitor AGA. In vivo, when B16 cells were mixed with 30% TK+ B16 cells, significantly reduced tumor weight and volume were observed after combinational treatment with resveratrol plus GCV as compared with GCV or resveratrol treatment alone. Conclusions Resveratrol could synergistically enhance the killing effect of TK/GCV suicide gene system in melanoma B16 cells and transplanted melanoma. It might be a promising adjuvant of TK/GCV therapy.