Resveratrol protects H9c2 cells against hypoxia-induced apoptosis through miR-30d-5p/SIRT1/NF-κB axis

Previous studies have demonstrated the cardioprotective role of resveratrol (Res). However, the underlyingmolecular mechanisms involved in the protective role of Res are still largely unknown. H9c2 cells weredistributed into five groups: normal condition (Control), DMSO, 20 mMRes (dissolved with DMSO), hypoxia(Hyp), and Res?Hyp. Cell apoptosis was evaluated using flow cytometry and protein analysis of cleavedcaspase 3 (cle-caspase 3). qRT-PCR assay was performed to measure the expression of microRNA-30d-5p(miR-30d-5p). MTT assay was performed to evaluate the cell proliferation. The relationship between miR-30d5p and silent information regulator 1 (SIRT1) was confirmed by luciferase reporter, RNA immunoprecipitation(RIP), and western blot assays. Western blot was performed to analyze NF-jB/p65 and I-jBa expressions. Ourdata showed that hypoxia enhanced apoptosis and NF-jB signaling pathway, which was alleviated by Restreatment. Hypoxia increased the expression of miR-30d-5p while decreased the SIRT1expression, which wasalso attenuated by Res treatment. Furthermore, miR-30d-5p depletion inhibited the proliferation, reducedapoptosis and decreased the expression of cle-caspase 3 in H9c2 cells with hypoxia treatment. Luciferasereporter, RIP, and western blot assays further confirmed that miR-30d-5p negatively regulated the expression ofSIRT1. Interestingly, the rescue-of-function experiments further indicated that knockdown of SIRT1 attenuatedthe effect of miR-30d-5p depletion on proliferation, apoptosis NF-jB signaling pathway inH9c2 cells withhypoxia treatment. In addition, the suppression of NF-jB signaling pathway increased cell viability whiledecreased cell apoptosis in hypoxia-mediatedH9c2 cells. Our data suggested Res mayprotectH9c2 cells againsthypoxia-induced apoptosis through miR-30d-5p/SIRT1/NF-jB axis

Research progress on the role of miR-30 family in the genesis and development of gastric cancer / 中国肿瘤生物治疗杂志

@#[摘 要] 胃癌（gastric carcinoma，GC）是最常见的消化系统恶性肿瘤之一，其病因及发病机制等都尚未明了，诊断、治疗及预后仍然面临着严峻的问题。微小RNA（microRNA，miRNA）作为一类重要的基因表达调节剂，能够与其相对应的靶基因相结合，从而影响基因表达，以此在GC发生发展中发挥重要的作用。miR-30家族中5个高度保守且成熟的成员——miR-30a、miR-30b、miR-30c、miR-30d和miR-30e位于不同的染色体或邻近位点，这些miRNA在5'末端附近有一个共同序列，3'末端附近具有不同的补偿序列。通过靶向各自相对应的基因影响着GC细胞的增殖、转移、侵袭和对化学药物的耐药性。miR-30家族在GC中发挥着重要的抑癌作用。本文就近年来miR-30家族中5个成员在GC发生发展中作用的研究进展作一综述。

The Effect of Antisense miR-30d on Invasion and Migration in Breast Cancer Cells / 天津医药

Objective To investigate the expression of miR-30d in breast cancer tissues and demonstrate the regula-tive effects of miR-30d ASO on the invasion and migration of breast cancer cells in vitro. Methods The expression levels of miR-30d in 108 breast cancer tissues and their adjacent tissues were detected by real-time quantitative PCR method. Af-ter transfection with miR-30d ASO, the biological effects of miR-30d on in breast cancer cells was measured by transwell as-say and wound healing assay. The expressions of matrix metalloproteinase (MMP)-2 and MMP-9 were analyzed by Western blot assay. Results The expression level of miR-30d was found to be over-expressed in breast cancer tissues(P<0.05). Compared with control group and nonsense interference group, the miR-30d expression was significantly decreased in breast cancer cells(transfection with miR-30d ASO). Results of transwell and wound healing assay showed that the invasion and mi-gration ability decreased significantly after transfection with miR-30d ASO, and expressions of MMP-2 and MMP-9 were down-regulated (P<0.05). Conclusion miR-30d was over-expressed in human breast cancer. The invasion and migration of breast cancer cells can be effectively inhibited by decreasing the expression of miR-30d. miR-30d may become a new tar-get for the regulation of invasion and migration in breast cancer.