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作 者:袁铭杰 刘天一[1] 陈亮[1] 王万晨 刘驰 毕波[1] YUAN MingJie;LIU Tianyi;CHEN Liang;WANG Wanchen;LIU Chi;BI Bo(Department of Plastic and Reconstructive Surgery,Fudan University Affiliated Hua Dong Hospital,Shanghai 200040,China)
机构地区:[1]复旦大学附属华东医院整形美容外科,上海200040
出 处:《中国麻风皮肤病杂志》2020年第12期712-717,共6页China Journal of Leprosy and Skin Diseases
基 金:上海市科委医学引导项目(西医)(编号:19411962300);上海市卫生健康委员会科研课题(编号:201940400)。
摘 要:目的:明确大黄素对黑素瘤B16F10细胞增殖、迁移及凋亡的影响。方法:10、20、40、60、80μmol/L的大黄素作用于B16F10细胞24 h后,Cell Counting Kit-8(CCK-8)法检测细胞增殖;倒置显微镜下观察细胞形态及数目变化;细胞划痕实验和Transwell迁移实验分别检测各组细胞划痕愈合率和细胞迁移数目;流式细胞术检测细胞周期;TUNEL法检测细胞凋亡情况;Western blot测定凋亡相关蛋白caspase-3的表达。结果:与空白对照组比较,不同浓度(10、20、40、60、80μmol/L)的大黄素均可抑制B16F10细胞的增殖(P<0.05),其抑制率呈浓度依赖和时间依赖关系,并且将细胞阻滞于G2/M期。在显微镜观察下细胞呈现出凋亡状态,不同浓度的大黄素组凋亡相关蛋白caspase-3的表达水平升高。结论:大黄素可有效抑制黑素瘤B16F10细胞增殖,抑制细胞迁移能力,并能促进细胞凋亡。Objective:To determine the effects of emodin on proliferation,migration and apoptosis of the melanoma cell line B16F10.Methods:The proliferation of B16F10 cells was detected by Cell Counting Kit-8(CCK-8)after treated with 10,20,40,60 and 80μmol/L emodin for 24 h.The cell morphology and number were observed under inverted microscope.The scratch healing rate and cell migration ability were detected by cell scratch test and Transwell migration assay.The cell cycle was detected with flow cytometry.The apoptosis of B16F10 cells was detected by TUNEL method.The expression level of casepase-3 was measured by Western blot.Results:Compared with the blank control group,emodin groups with different concentrations(10,20,40,60 and 80μmol/L)inhibited the proliferation of B16F10 cells and the inhibition rate showed a concentration-and time-dependent manner.The cell cycle was arrested at G2/M phase.The cell apoptosis was observed under microscope.The expression level of caspase-3 in the emodin groups was higher that in the control group.Conclusion:Emodin can effectively inhibit the proliferation and migration of B16F10 cells and induce cell apoptosis.
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