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长链非编码 RNA DUXAP8 通过靶向 miR-590-5p 调控卵巢癌细胞的增殖和凋亡。

Long noncoding RNA DUXAP8 regulates proliferation and apoptosis of ovarian cancer cells via targeting miR-590-5p.

机构信息

Department of Women's Healthcare, Jinan Maternity and Child Care Hospital, Jinan, 250002, China.

Department of Pathology, Jinan Maternity and Child Care Hospital, No.2 Xiaojing 3rd Jianguo Road, Jinan, 250002, China.

出版信息

Hum Cell. 2020 Oct;33(4):1240-1251. doi: 10.1007/s13577-020-00398-8. Epub 2020 Aug 4.

Abstract

The aim of this study is to investigate the effect of lncRNA DUXAP8 on proliferation and apoptosis of ovarian cancer cells, and to explore its potential mechanism. DUXAP8 interfering and overexpressing cell lines were constructed and the cell proliferation and apoptosis were tested. Hematoxylin-eosin, TdT-mediated dUTP nick end labeling, and immunohistochemistry were used to detect the effect of DUXAP8 on the ability of tumor formation. Quantitative real-time polymerase chain reaction and western blot were used to detect the mRNA and protein expression of miR-590-5p and YAP1, respectively. Dual luciferase assay was used to determine the target relationship between DUXAP8, miR-590-5p, and YAP1. DUXAP8 interference and miR-590-5p down-regulated cell lines were further constructed. Compared with normal ovarian cells, the expression of DUXAP8 in ovarian cancer cells was significantly increased, while the expression of miR-590-5p was decreased (p < 0.05). After DUXAP8 interference, cell proliferation and colony formation were decreased, and apoptosis was increased. The results of in vivo experiment are consistent with the in vitro experiments. The expression of miR-590-5p was up-regulated and the expression of YAP1 was decreased after DUXAP8 interference. Moreover, miR590-5p inhibitor can attenuate the effect of DUXAP8 interference on ovarian cancer cells. Taken together, lncRNA DUXAP8 can regulate the proliferation and apoptosis of ovarian cancer cells, and its mechanism may be related to the regulation of YAP1 gene by targeting miR-590-5p.

摘要

本研究旨在探讨长链非编码 RNA DUXAP8 对卵巢癌细胞增殖和凋亡的影响,并探讨其潜在机制。构建了 DUXAP8 干扰和过表达细胞系,并检测了细胞增殖和凋亡。苏木精-伊红、TdT 介导的 dUTP 缺口末端标记和免疫组织化学用于检测 DUXAP8 对肿瘤形成能力的影响。实时定量聚合酶链反应和蛋白质印迹分别用于检测 miR-590-5p 和 YAP1 的 mRNA 和蛋白表达。双荧光素酶报告基因实验用于确定 DUXAP8、miR-590-5p 和 YAP1 之间的靶关系。进一步构建了 DUXAP8 干扰和 miR-590-5p 下调细胞系。与正常卵巢细胞相比,卵巢癌细胞中 DUXAP8 的表达明显增加,而 miR-590-5p 的表达则降低(p<0.05)。干扰 DUXAP8 后,细胞增殖和集落形成减少,凋亡增加。体外实验结果与体内实验结果一致。干扰 DUXAP8 后 miR-590-5p 的表达上调,YAP1 的表达下调。此外,miR590-5p 抑制剂可减弱 DUXAP8 干扰对卵巢癌细胞的影响。总之,lncRNA DUXAP8 可以调节卵巢癌细胞的增殖和凋亡,其机制可能与通过靶向 miR-590-5p 调节 YAP1 基因有关。

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