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miR-28-5p 介导姜黄素对人弥漫大 B 细胞淋巴瘤细胞的抗增殖和促凋亡作用。

MiR-28-5p mediates the anti-proliferative and pro-apoptotic effects of curcumin on human diffuse large B-cell lymphoma cells.

机构信息

Department of Pediatrics, People's Hospital of Shijiazhuang City, Shijiazhuang, China.

Department of Rehabilitation, The Second Hospital of Hebei Medical University, Shijiazhuang, China.

出版信息

J Int Med Res. 2020 Jul;48(7):300060520943792. doi: 10.1177/0300060520943792.

Abstract

OBJECTIVE

To investigate the anti-proliferative and pro-apoptotic effects of curcumin on diffuse large B-cell lymphoma (DLBCL) cells and explore the mechanism.

METHODS

OCI-LY7 cells were treated with curcumin (2.5, 5, 10, 20, and 40 μM) for 24, 48, or 72 hours. Cell viability and apoptosis were determined using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5 diphenyl tetrazolium bromide assay and TdT-mediated dUTP nick-end labeling staining, respectively. MiR-28-5p expression was detected via qRT-PCR. The binding site of miR-28-5p was predicted using online databases and verified using the dual-luciferase reporter assay. MiR-28-5p overexpression and inhibition were achieved via transfection with an miR-28-5p mimic and inhibitor, respectively.

RESULTS

Curcumin decreased the viability of OCI-LY7 cells in a concentration- and time-dependent manner, and these effects were attenuated by miR-28-5p inhibition. MiR-28-5p expression was upregulated by curcumin. Curcumin increased the numbers of apoptotic cells and upregulated cleaved caspase-3 expression, and these effects were attenuated by miR-28-5p inhibition. The dual-luciferase reporter assay confirmed that miR-28-5p directly targets the 3'-untranslated region of BECN1. Curcumin downregulated BECN1 and microtubule-associated protein 1 light chain 3 beta-II/I expression and upregulated p62 expression.

CONCLUSIONS

Our results described the curcumin exerted anti-proliferative and pro-apoptotic effects on OCI-LY7 cells through a mechanism potentially involving miR-28-5p.

摘要

目的

研究姜黄素对弥漫性大 B 细胞淋巴瘤(DLBCL)细胞的抗增殖和促凋亡作用,并探讨其机制。

方法

用姜黄素(2.5、5、10、20 和 40 μM)处理 OCI-LY7 细胞 24、48 或 72 小时。用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)比色法和末端转移酶介导的 dUTP 缺口末端标记染色法分别检测细胞活力和细胞凋亡。用 qRT-PCR 检测 miR-28-5p 的表达。利用在线数据库预测 miR-28-5p 的结合位点,并通过双荧光素酶报告基因检测进行验证。通过转染 miR-28-5p 模拟物和抑制剂分别实现 miR-28-5p 的过表达和抑制。

结果

姜黄素呈浓度和时间依赖性地降低 OCI-LY7 细胞的活力,而 miR-28-5p 抑制则减弱了这些作用。姜黄素上调 miR-28-5p 的表达。姜黄素增加凋亡细胞数量并上调 cleaved caspase-3 表达,而 miR-28-5p 抑制则减弱了这些作用。双荧光素酶报告基因检测证实 miR-28-5p 可直接靶向 BECN1 的 3'-非翻译区。姜黄素下调 BECN1 和微管相关蛋白 1 轻链 3β-II/I 的表达,并上调 p62 的表达。

结论

我们的结果描述了姜黄素通过一种可能涉及 miR-28-5p 的机制对 OCI-LY7 细胞发挥抗增殖和促凋亡作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2fa/7388109/abf23a29c25d/10.1177_0300060520943792-fig1.jpg

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