桑黄水提物诱导黑色素瘤细胞系B16-F10 G_(0)/G_(1)期阻滞的研究  

Study on the Induction of G_(0)/G_(1) Phase Arrest in Melanoma Cell Line B16-F10 by Aqueous Extracts of Sanghuang

作　　者：霍进喜 李有贵[1] 孙雨晴[1] 潘美良 马焕艳 梁菲 钟石[1] Huo Jinxi;LiYougui;Sun Yuqing;Pan Meiliang;Ma Huanyan;Liang Fei;Zhong Shi(Institute of Sericulture and Tea,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China;Zhejiang Provincial Center for Agricultural Technology Extension,Hangzhou 310020,China;School of Basic Medical Sciences&Forensic Medicine,Hangzhou Medical College,Hangzhou 310059,China)

机构地区：[1]浙江省农业科学院蚕桑与茶叶研究所,杭州310021 [2]浙江省农业技术推广中心,杭州310020 [3]杭州医学院基础医学与法医学院,杭州310059

出　　处：《蚕业科学》2023年第6期544-550,共7页ACTA SERICOLOGICA SINICA

基　　金：浙江省自然科学基金项目(LQ21C150002);浙江省重点研发计划项目(2018C02003);杭州医学院院所专项(YS2021011)。

摘　　要：通过噻唑蓝(MTT)比色法、流式细胞分析、转录组测序和蛋白印迹等手段,探究桑黄水提物(SH)对黑色素瘤细胞系B16-F10的抑制作用及分子作用机制。MTT实验结果显示,SH能够抑制B16-F10细胞系增殖,且呈剂量依赖性。流式细胞分析发现,SH能够诱导细胞G_(0)/G_(1)期阻滞,但对细胞凋亡无明显影响。RNA-seq、qRT-PCR和Western blot分析发现,SH主要通过上调p53信号通路中p21的表达,进而抑制细胞周期通路中CyclinD3、CDK2和CDK6的表达来影响细胞周期阻滞。研究结果证明SH对黑色素瘤细胞增殖具有显著的抑制作用,p21-CyclinD3-CDKs信号通路介导的G_(0)/G_(1)期阻滞是其作用机制之一。The inhibitory effects and molecular mechanism of aqueous extracts of Sanghuang(SH) on melanoma cell line B16-F10 were investigated by thiazole blue(MTT) colorimetry, flow cytometry, transcriptome sequencing and Western blot. The result of MTT assay showed that SH could inhibit the proliferation of B16-F10 cell lines in a dose-dependent manner. Flow cytometry analysis found that SH could induce B16-F10 cell cycle arrest at G_(0)/G_(1) phase, whereas there were no obvious effects on apoptosis. RNA-seq, qRT-PCR and Western blot analysis showed that SH mainly affected cell cycle arrest by up-regulating the expression of p21 in the p53 signaling pathway, thereby inhibiting the expression of CyclinD3, CDK2 and CDK6 in the cell cycle pathway. The research results demonstrate that SH has significant inhibitory effects on the proliferation of melanoma cells, and the G_(0)/G_(1) phase arrest mediated by the p21-CyclinD3-CDKs signaling pathway is one of the mechanisms of action.

关 键 词：桑黄 水提物 G_0/G_1期阻滞 黑色素细胞瘤 

分 类 号：R285.5[医药卫生—中药学] S886.9[医药卫生—中医学]