基础医学与临床2025，Vol.45Issue(1) ：7-11.DOI:10.16352/j.issn.1001-6325.2025.01.0007

褪黑素促进小鼠黑色素瘤细胞系B16-F10失巢凋亡

Melatonin promotes anoikis of mouse melanoma cell line B16-F10

甘雨灵 李廷栋 刘利兵 周英芬 潘东升
基础医学与临床2025，Vol.45Issue(1) ：7-11.DOI:10.16352/j.issn.1001-6325.2025.01.0007
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  • NSTL
  • 万方数据

褪黑素促进小鼠黑色素瘤细胞系B16-F10失巢凋亡

Melatonin promotes anoikis of mouse melanoma cell line B16-F10

甘雨灵 1李廷栋 1刘利兵 1周英芬 1潘东升1
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作者信息

  • 1. 甘肃省肿瘤医院 骨与软组织肿瘤一科,甘肃 兰州 730000
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摘要

目的 探讨褪黑素对黑色素瘤细胞系失巢凋亡的作用及机制.方法 通过CCK-8 法筛选褪黑素抑制黑色素瘤细胞系B16-F10 的最佳药物浓度;将细胞分为 4 组:抗失巢凋亡模型组、TrkB激活剂组、褪黑素组、褪黑素+TrkB激活剂组;细胞活性/细胞毒性试剂盒(Calcein AM/EthD-1)检测黑色素瘤细胞失巢凋亡;荧光探针DCFH-DA进行活性氧检测;Western blot检测各组Nrf2 蛋白和TrkB蛋白的表达.结果 褪黑素可以明显抑制黑色素瘤细胞的增殖,呈时间与剂量依赖性,半数抑制浓度IC50为 1×10-7μmol/L,其抑制作用与诱导黑色素瘤细胞的失巢凋亡有关.褪黑素可以上调细胞活性氧生成(P<0.05),而加入TrkB激活剂可以拮抗此作用.褪黑素可降低黑色素瘤细胞中Nrf2 蛋白和TrkB蛋白表达(P<0.05),而加入TrkB激活剂可以抑制褪黑素对Nrf2 蛋白和TrkB蛋白表达的影响(P<0.05).结论 褪黑素可通过诱导细胞失巢凋亡来抑制黑色素瘤细胞系B16-F10 的增殖.

Abstract

Objective To investigate the effect and mechanism of melatonin on the anoikis of melanoma cells.Methods The drug concentration of melatonin inhibiting melanoma cell line B16-F10 was optimized based on the effect on CCK-8 assay.An anti-anoikis of melanoma cell model was developed and divided it into four groups:The blank control group,the TrkB activator group,the melatonin group and the melatonin+TrkB activator group.Calce-in AM/EthD-1 fluorescence double staining was used to detect the anoikis of melanoma cells.Reactive oxygen spe-cies were detected using the fluorescent probe DCFH-DA.Western blot was used to detect the expression of Nrf2 protein and TrkB protein in each group.Results Melatonin significantly inhibited the proliferation of melanoma cells in a time-and dose-dependent manner with IC50 of 1×10-7 μmol/L.Its inhibitory effect was found to be related to in-duction of anoikis of melanoma cells.Melatonin could upregulate the generation of cellular reactive oxygen species(P<0.05),while addition of TrkB activator antagonized this effect.Melatonin could reduce the expression of Nrf2 protein and TrkB protein in melanoma cells(P<0.05),and the addition of TrkB activator could inhibite the effect of melatonin on the expression of Nrf2 protein and TrkB protein(P<0.05).Conclusions Melatonin can inhibit the pro-liferation of melanoma cell line B16-F10 through the mechanism of inducing anoikis.

关键词

黑色素瘤/褪黑素/失巢凋亡/活性氧

Key words

melanoma/melatonin/anoikis/reactive oxygen species

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出版年

2025
基础医学与临床
北京生理科学会

基础医学与临床

影响因子：0.669
ISSN：1001-6325
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