Aloe-Emodin Induces Breast Tumor Cell Apoptosis through Upregulation of miR-15a/miR-16-1 That Suppresses BCL2
Aloe-Emodin Induces Breast Tumor Cell Apoptosis through Upregulation of miR-15a/miR-16-1 That Suppresses BCL2
复制标题
芦荟大黄素通过上调抑制 BCL2 的 miR-15a/miR-16-1 诱导乳腺肿瘤细胞凋亡
DOI：
10.1155/2020/5108298
复制
发表时间：
2020-03-04
影响因子：
--
通讯作者：
Ma, Min
中科院分区：
文献类型：
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作者：
Jiang, Xuefeng;Liu, Yusheng;Ma, Min
目的芦荟大黄素（Aloe-emodin，AE）是从芦荟（Aloe vera）和大黄（Palmatum rheumarrhythm）中提取的天然化合物，对多种肿瘤具有抗癌活性。Bcl-2家族是细胞死亡或细胞存活的主要调节因子。本研究描述了AE对乳腺肿瘤（BT）细胞增殖的影响。方法采用AE处理MCF-10A、MCF-10AT、MCF-7和MDA-MB-231细胞株。CCK-8和流式细胞仪检测细胞增殖和凋亡。蛋白质水平通过蛋白质印迹法测量。RT-PCR检测mRNA和miRNA水平。应用生物信息学方法筛选与mRNA 3′-UTR结合的miRNAs。结果AE选择性抑制MCF-10AT和MCF-7细胞增殖并诱导其凋亡，但对MCF-10A和MDA-MB-231细胞无明显抑制作用。AE剂量依赖性地降低MCF-10AT和MCF-7细胞中Bcl-2和Bcl-xl的蛋白表达，而增加Bax蛋白表达。AE处理后Bcl-xl和Bax mRNA水平发生变化，与蛋白表达结果一致。然而，两种细胞系中的Bcl-2 mRNA水平均未受到影响，这表明AE可能通过miRNA调节Bcl-2的蛋白翻译。在所有与Bcl-2的3′-UTR结合的候选miRNAs中，miR-15 a和miR-16-1被AE剂量依赖性地下调。抑制miR-15 a/16-1可消除AE对MCF-10AT和MCF-7细胞生长的抑制作用，并逆转AE对Bcl-2蛋白水平的下调。结论本研究为AE通过上调miR-15 a/miR-16-1抑制BCL 2诱导BT细胞凋亡提供了重要依据。
Purpose Aloe-emodin (AE) is a natural compound derived from aloe vera and palmatum rhubarb and shows anticancer activities in various cancers. Bcl-2 family is the main regulator of cell death or cell survival. This study describes the effects of AE on proliferation of breast tumor (BT) cells. Methods MCF-10A, MCF-10AT, MCF-7, and MDA-MB-231 cell lines were exposed to AE. Cell proliferation and apoptosis were assessed by CCK-8 and flow cytometry. Protein levels were measured by Western blotting. The levels of mRNA and miRNA were examined by RT-PCR. Bioinformatics was applied to screen miRNAs that bind to 3′-UTR of mRNA. Results The results showed that AE selective activity inhibited the proliferation and induced apoptosis of MCF-10AT and MCF-7 cells but exhibited no significant inhibition in MCF10A and MDA-MB-231 cells. Mechanistically, AE dose-dependently decreased the protein expression of Bcl-2 and Bcl-xl, while it increased Bax protein expression in MCF-10AT and MCF-7 cells. The levels of Bcl-xl and Bax mRNA were altered by AE treatment, which was consistent with the protein expression results. However, Bcl-2 mRNA levels were not affected in either cell line, suggesting that AE may modulate the protein translation of Bcl-2 through miRNAs. In all candidate miRNAs that bind to 3′-UTR of Bcl-2, miR-15a and miR-16-1 were dose-dependently downregulated by AE. Moreover, inhibition of miR-15a/16-1 could eliminate the inhibition of MCF-10AT and MCF-7 cells growth by AE and could reverse the downregulation of AE-induced Bcl-2 protein level. Conclusion Our research provides an important basis that AE induces BT cell apoptosis through upregulation of miR-15a/miR-16-1 that suppresses BCL2.