大黄素通过调节miR-582-3p影响MAP3K1活性抑制乳腺癌细胞增殖并诱导凋亡  

Emodin inhibits proliferation and induces apoptosis of breast cancer cells by down-regulating miR-582-3p to suppress MAP3K1

作　　者：范敏慧 帅仁亚 李晴宇[2] FAN Min-hui;SHUAI Ren-ya;Li Qing-yu(Department of Pharmacy,Hangzhou Cancer Hospital,Hangzhou 310002,China;Department of Pharmacy,Wushan Branch of Hangzhou First People's Hospital,Hangzhou 310006,China)

机构地区：[1]杭州市肿瘤医院药学部,杭州310002 [2]杭州市第一人民医院药学部,杭州310006

出　　处：《浙江中西医结合杂志》2024年第5期407-414,共8页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省中医药科技计划项目(No.20222B286)。

摘　　要：目的 探讨大黄素(emodin,EMO)通过microRNA(miR)-582-3p/MAP3K1对乳腺癌细胞增殖、凋亡的作用及分子机制。方法 培养人乳腺癌细胞株MCF-7、MDA-MB-231以及人乳腺细胞系Hs578Bst,使用不同浓度的EMO处理细胞,细胞根据培养条件分为对照组、EMO(10μM)组、NC mimics组、NC mimics+EMO组、miR-582-3p mimics组和miR-582-3p mimics+EMO组,检测EMO及miR-582-3p异常表达对乳腺癌细胞增殖及凋亡的影响。荧光定量(qRT-PCR)检测miR-582-3p及MAP3K1的表达。MTT和EdU实验检测乳腺癌细胞增殖,流式细胞术和TUNEL实验检测细胞凋亡及细胞周期。蛋白印迹(Western blot)检测增殖标志蛋白Ki-67及凋亡标志蛋白Bcl-2、Bax和Caspase-3在癌细胞中的表达水平。结果 EMO可以抑制乳腺癌细胞MDA-MB-231及MCF-7的增殖能力[(17.65±0.96)%比(28.63±2.16)%,(15.02±1.71)%比(28.22±1.89)%,P<0.05]、调节细胞周期并促进细胞凋亡[(23.67±1.25)%比(5.73±0.84)%,(27.33±1.73)%比(5.17±0.85)%,P<0.05]。Western blot结果显示,EMO可抑制细胞中Ki-67及Bcl-2的表达并促进Caspase-3(cleaved)及Bax的表达(P<0.05)。EMO处理乳腺癌细胞后miR-582-3p的表达显著下调[(0.45±0.09)比(1.00±0.10),(0.37±0.05)比(1.00±0.11),P<0.01],而过表达miR-582-3p会逆转EMO对乳腺癌细胞生物学活性的影响(P<0.05)。EMO还会通过miR-583-3p调节癌细胞中MAP3K1的表达(P<0.05)。结论 EMO可能通过调节miR-582-3p影响MAP3K1活性抑制乳腺癌细胞增殖并诱导凋亡。Objective To investigate the effects and molecular mechanisms of emodin(EMO)on proliferation and apoptosis of breast cancer cells via microRNA(miR)-582-3p/MAP3K1.Methods Human breast cancer cell lines,MCF-7 and MDA-MB-231,as well as human breast cell strain Hs578Bst,were cultured and treated with different concentrations of EMO.According to different culture conditions,the cells were divided into control,EMO(10μM),NC mimics,NC mimics+EMO,miR-582-3p mimics,and miR-582-3p mimics+EMO groups.The effects of EMO and abnormal expression of miR-582-3p on the proliferation and apoptosis of breast cancer cells were assessed.qRT-PCR was used to detect the expression of miR-582-3p and MAP3K1.MTT and EdU assays were carried out to detect breast cancer cell proliferation.Flow cytometry and TUNEL assays were conducted to assess cell apoptosis and cell cycle.Western blot was employed for detecting the expression levels of proliferation marker protein Ki-67 and apoptosis marker proteins Bcl-2,Bax,and Caspase-3 in cancer cells.Results Compared to the control,EMO inhibited the proliferation of MDA-MB-231 and MCF-7[(17.65±0.96)vs.(28.63±2.16)%,(15.02±1.71)vs.(28.22±1.89)%,P<0.05],regulated cell cycle,and promoted cell apoptosis[(23.67±1.25)vs.(5.73±0.84)%,(27.33±1.73)vs.(5.17±0.85)%,P<0.05].Western blot results showed that EMO inhibited the expression of Ki-67 and Bcl-2 while promoting the expression of Caspase-3(cleaved)and Bax(P<0.05)in cells.The expression of miR-582-3p was significantly down-regulated after breast cancer cells were treated with EMO[(0.45±0.09)vs.(1.00±0.10)%,(0.37±0.05)vs.(1.00±0.11)%,P<0.01],while overexpression of miR-582-3p reversed the effect of EMO on the biological activity of breast cancer cells(P<0.05).Additionally,EMO regulated the expression of MAP3K1 in cancer cells through miR-583-3p(P<0.05).Conclusion EMO inhibits proliferation and induces apoptosis of breast cancer cells through down-regulation of miR-582-3p to suppress MAP3K1.

关 键 词：大黄素 乳腺癌细胞 miR-582-3p MAP3K1 

分 类 号：R285[医药卫生—中药学]