Tagitinin A抑制黑色素瘤细胞SK-Mel-28的机制

The mechanism of Tagitinin A anti-melanoma SK-Mel-28 cells

摘要

目的 探究化合物Tagitinin A对黑色素瘤细胞的抑制作用及其分子机制。方法 本研究以黑色素瘤SK-Mel-28细胞为模型,采用不同浓度的Tagitinin A（1.25～20 μmol/L）处理SK-Mel-28细胞。通过噻唑蓝（MTT）和平板克隆形成实验检测细胞的存活率和克隆形成能力；采用流式细胞术检测细胞周期变化和凋亡；划痕实验检测细胞迁移能力变化；使用DCFH-DA标记的荧光探针检测经Tagitinin A处理后的黑色素瘤细胞内活性氧（ROS）的水平；最后,通过免疫印迹（WB）实验检测细胞内质网应激和凋亡相关信号通路的蛋白表达。结果 与对照组相比,Tagitinin A处理的SK-Mel-28细胞生长和平板克隆形成能力显著降低,细胞迁移能力明显减弱,且细胞周期阻滞在G₂/M期。此外,Tagitinin A诱导细胞内活性氧增加,上调ER stress相关蛋白如PERK、CHOP和促凋亡蛋白BAX的表达,且抑制抗凋亡蛋白Bcl-2表达。结论 Tagitinin A可能通过诱导ROS生成,激活内质网应激信号通路并诱导细胞凋亡,进而抑制黑色素瘤SK-Mel-28细胞增殖与转移。

Abstract

Objective To explore inhibitory effect on melanoma cells of compound Tagitinin A and the molecular mechanism of its anticancer activity. Methods In this study, SK-Mel-28 melanoma cells were used as a model. After treatment with different concentrations of Tagitinin A (1.25-20 μmol/L), the cell viability and colony formation ability were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and colony formation assay, and the changes of cell cycle and apoptosis were detected by flow cytometry. We used wound healing assay to detect the changes in cell migration ability. DCFH-DA labeled fluorescent probe was used to detect the level of reactive oxygen species (ROS) in melanoma cells, and measurement of the protein expression of endoplasmic reticulum stress and apoptosis-related signaling pathways was performed with Western blot. Results Compared with the control group, Tagitinin A significantly inhibited the growth and colony formation of SK-Mel-28 cells, reduced cell migration, and arrested the cell cycle at G₂/M phase. In addition, Tagitinin A increased intracellular reactive oxygen species, increased the expression of ER stress related proteins PERK and CHOP, increased the pro-apoptotic protein BAX, and decreased the anti-apoptotic protein Bcl-2. Conclusion Tagitinin A may suppress the proliferation and migration of melanoma SK-Mel-28 cells by inducing the accumulation of ROS, activating ER stress signaling pathway and promoting apoptosis.

关键词

化合物Tagitinin A / 黑色素瘤 / 增殖 / 凋亡 / endoplasmic reticulum stress信号通路

Key words

compound Tagitinin A / melanoma / proliferation / apoptosis / endoplasmic reticulum stress signaling pathway

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基金资助

贵州省科技计划项目(黔科合基础MS(2026)932)

遵义医科大学博士启动基金资助项目(ZMUDRF2025-37)

遵义医科大学博士启动基金资助项目(ZMUDRF2025-34)