摘要
目的探究白藜芦醇(resveratrol,RES)抑制葡萄膜黑色素瘤细胞(choroidal melanoma cells,MUM2B)自噬并促进其凋亡的调控作用及机制。方法将MUM2B分为对照组、实验组,分别用不同浓度的RES(0、10、20、40、60、80μmol·L-1)处理细胞。划痕实验检测细胞迁移率,CCK-8检测细胞活力,流式细胞术检测细胞凋亡,通过qRT-PCR和Western blot检测miR-512-3P、DUSP1以及其他目的蛋白的表达,RIP实验验证miR-512-3P靶向沉默DUSP1,使用miR-512-3P的mimic,体外评估miR-512-3P的生物学意义。结果RES抑制了MUM2B细胞的增殖与侵袭,减弱细胞自噬,促进细胞凋亡;MUM2B细胞p-ERK、p-MTOR蛋白表达明显增加,miR-512-3P表达增加;过表达miR-512-3P靶向抑制了DUSP1的表达,并促进了MUM2B细胞的凋亡。结论RES通过促进miR-512-3P的表达靶向抑制DUSP1,从而抑制MUM2B的自噬并促进其凋亡。
Aim To investigate the regulatory role and mechanism of resveratrol in inhibiting autophagy and promoting apoptosis in choroidal melanoma cells.Methods Choroidal melanoma cells(MUM2B)were divided into control and experimental groups,and treated with different concentrations of resveratrol(0,10,20,40,60,80μmol·L-1).Cell migration was detected by scratch assay,cell viability was detected by CCK-8,cell apoptosis was detected by flow cytometry,and the expressions of miR-512-3P,DUSP1,and other target proteins were detected by qRT-PCR and Western blot.miR-512-3P was verified to be targeted to the silencing of DUSP1 by RIP.The biological significance of miR-512-3P was assessed in vitro using miR-512-3P mimic.Results Resveratrol inhibited the proliferation and invasion of MUM2B cells,attenuated cellular autophagy,and promoted apoptosis.In addition,cellular p-ERK and p-MTOR protein expression significantly increased,and miR-512-3P expression increased.Overexpression of miR-512-3P targeted and inhibited DUSP1 expression and promoted apoptosis in MUM2B cells.Conclusion Resveratrol targets and inhibits DUSP1 by promoting miR-512-3P expression,thereby inhibiting autophagy and promoting apoptosis in uveal melanoma cells.
作者
孙正杨
刘楠楠
范学菲
陈苏环
陈唔奇
陈广祎
邵玉宝
陈晓宇
SUN Zheng-yang;LIU Nan-nan;FAN Xue-fei;CHEN Su-huan;CHEN Wu-qi;CHEN Guang-yi;SHAO Yu-bao;CHEN Xiao-yu(Dept of Histology and Embryology,Anhui Medical University,Hefei 230032,China;Dept of Ophthalmology,Hospital of University of Science and Technology of China,Hefei 230002,China;Dept of Clinical Medicine,Anhui Medical University,Hefei 230032,China;Experimental Center of Morphology,Anhui Medical University,Hefei 230032,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2024年第2期292-298,共7页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81373421)
安徽医科大学“早期接触科研”项目(No 2022-ZQKY-19)。
