杨梅枝条醇提物对A375细胞增殖和凋亡的影响及其分子机制  

Effect of alcohol extracts from Chinese bayberry branch on proliferation and apoptosis of A375 cells and its molecular mechanism

作　　者：郑园园 俞浙萍 张淑文[2] 李有贵[3] 孙鹂[2] 郑锡良[2] 戚行江 ZHENG Yuanyuan;YU Zheping;ZHANG Shuwen;LI Yougui;SUN Li;ZHENG Xiliang;QI Xingjiang(College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, Zhejiang, China;Institute of Horticulture, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;Institute of Sericultural and Tea, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China)

机构地区：[1]浙江师范大学化学与生命科学学院,浙江金华321004 [2]浙江省农业科学院园艺研究所,浙江杭州310021 [3]浙江省农业科学院蚕桑与茶叶研究所,浙江杭州310021

出　　处：《浙江农业学报》2022年第5期974-983,共10页Acta Agriculturae Zhejiangensis

基　　金：浙江省农业(果品)新品种选育重大科技专项(2021C02066-2);浙江省重点研发计划(2021C02009)。

摘　　要：杨梅(Myrica rubra)具有较高的药用价值,利用杨梅枝条醇提物(Myrica rubra branch extract,MRBE)开展其对人恶性黑色素瘤A375细胞的作用研究。采用MTT法、PI流式细胞术、Annexin V-PI双染流式细胞术、实时荧光定量PCR(qRT-PCR)等方法,解析MRBE诱导A375细胞阻滞与凋亡的潜在作用机制。结果表明:MRBE对A375细胞的抑制率呈剂量依赖效应关系,抑制率最高达49.46%;流式细胞术结果表明,MRBE作用48 h可使A375细胞阻滞于S期并发生凋亡;qRT-PCR结果进一步表明,MRBE可能通过上调P21和下调CyclinA、CyclinD、CDK2、CDK4、CDK6、E2F1等基因的表达,诱导A375细胞阻滞于S期,同时下调Bcl-w和Raf基因的表达促使A375细胞凋亡。初步解析了MRBE对人恶性黑色素瘤A375细胞增殖和凋亡的作用机制,为利用MRBE研发抗黑色素瘤潜在药物提供科学依据,同时有助于提高杨梅枝条的药用附加值,拓宽杨梅产业链多元化发展。Chinese bayberry(Myrica rubra)has high medicinal values.This study was performed to evaluate the effect of alcohol extracts of Myrica rubra branch extract(MRBE)on human malignant melanoma cells A375.MTT assay,flow cytometry based on PI,Annexin V/PI double staining and quantitative reverse-transcription PCR(qRT-PCR)were used to dissect the potential mechanism of the arrest and apoptosis of A375 cells induced by MRBE.The results revealed that MRBE inhibited the proliferation of human A375 cells in a dose-dependent way,and the inhibition rate was up to 49.46%.Flow cytometry analyses suggested that 48 h treatment of MRBE induced both S-phase cell cycle arrest and apoptotic death in A375 cells.The results of qRT-PCR further showed that MRBE induced S phase arrest by promoting the upregulation of p21 and decreasing the expression of CyclinA,CyclinD,CDK2,CDK4,CDK6 and E2F1,and induced apoptosis by decreasing the expression of Bcl-w and Raf.This study revealed the action mechanism of MRBE on the induction of S-phase arrest and apoptosis of A375 cells,which could provide scientific evidence for the development of potential anti-melanom drugs by using MRBE,and help to improve the medicinal added value of Chinese bayberry branches and broaden the diversified development of its industry chain.

关 键 词：杨梅 枝条醇提物 人黑色素瘤A375细胞 细胞周期 细胞凋亡 

分 类 号：S567.1+9[农业科学—中草药栽培] R285[农业科学—作物学]