碘化丙啶
细胞凋亡
流式细胞术
分子生物学
细胞周期
免疫印迹
细胞生长
白藜芦醇
转染
化学
生物
细胞计数
染色
细胞
圆周率
膜联蛋白
细胞培养
程序性细胞死亡
生物化学
基因
遗传学
作者
Nan Zhou,Shuxing Cao,Zhenzhen Wang,Jianmin Luo,Xiaojun Liu,Yintao Shang,Lin Yang
出处
期刊:PubMed
日期:2023-08-01
卷期号:31 (4): 1093-1099
标识
DOI:10.19746/j.cnki.issn.1009-2137.2023.04.025
摘要
To investigate the effect of resveratrol (RSV) on the proliferation of multiple myeloma (MM) cells and its molecular mechanism.MM cells (MM1.S, RPMI-8226 and U266) were treated with different concentrations of RSV for 24-72 h. The effect of RSV on the proliferation of MM cells was detected by CCK-8 (cell counting kit-8) assay. RPMI-8226 cells were divided into RSV, miR-21 mimic, RSV+miR-21 mimic, miR-21 inhibitor and RSV+miR-21 inhibitor groups, and transfected with corresponding plasmids. The cell cycle distribution of each group was detected by flow cytometry with propidium iodide (PI) single staining. The cell apoptosis of each group was detected by AnnexinV-FITC/PE-PI double staining. The expression of miR-21 in MM cells treated with RSV and the expression of KLF5 mRNA in each group were detected by qRT-PCR. The expression of KLF5 protein in each group was detected by Western blot.RSV inhibited the proliferation and induced apoptosis of MM cells in a time- and dose-dependent manner. After the MM cells were treated with RSV, the number of cells in sub-G1 phase was increased, and that in G2/M phase was decreased. Moreover, RSV significantly downregulated the expression of miR-21 in MM cells, and the inhibitory effect of miR-21 mimic on KLF5 expression in MM cells was counteracted by RSV.RSV may inhibit the proliferation and induce apoptosis of MM cells by inhibiting miR-21 and up-regulating KLF5 expression.白藜芦醇对多发性骨髓瘤细胞的增殖抑制作用及相关机制研究.探讨白藜芦醇(resveratrol,RSV)对多发性骨髓瘤(MM)细胞增殖的影响及其分子机制。.不同浓度的RSV作用于MM细胞(MM1.S、RPMI-8226和U266)24-72 h,CCK-8法检测RSV对MM细胞增殖的影响;将RPMI-8226细胞分为RSV、miR-21 mimic、RSV+miR-21 mimic、miR-21 inhibitor和RSV+miR-21 inhibitor组,转染相应的质粒,碘化丙啶单染流式细胞术检测各组细胞的周期分布,AnnexinV-FITC/PE-PI双染法观察各组细胞的凋亡情况,qRT-PCR技术检测RSV处理后MM细胞中miR-21的表达水平和各组细胞中的KLF5 miRNA表达水平,Western blot技术检测各组细胞中的KLF5蛋白的表达水平。.RSV呈时间-剂量依赖性抑制MM细胞增殖,诱导细胞凋亡发生;RSV处理MM细胞后,亚G1期细胞增多,G2/M期细胞减少;同时RSV显著下调了MM细胞中miR-21 的表达,且给予RSV后抵消了miR-21 mimic对MM细胞中KLF5表达的抑制作用。.RSV通过抑制miR-21上调KLF5的表达水平,从而抑制MM细胞增殖并诱导其凋亡的发生。.
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