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环状 RNA RBM33 通过调节 miR-758-3p/PUM2 轴促进宫颈癌的进展。

Circular RNA RBM33 contributes to cervical cancer progression via modulation of the miR-758-3p/PUM2 axis.

机构信息

Department of Obstetrics & Gynaecology, the Affiliated Jintan Hospital of Jiangsu University, Changzhou, 213200, People's Republic of China.

Department of Obstetrics & Gynaecology, The Affiliated Jintan Hospital of Jiangsu University, No. 16, Nanmen Street, Jintan District, Changzhou City, Jiangsu Province, People's Republic of China.

出版信息

J Mol Histol. 2021 Apr;52(2):173-185. doi: 10.1007/s10735-020-09933-1. Epub 2021 Jan 5.

Abstract

Cervical cancer (CC) is a gynecological malignant tumor. Circular RNA (hsa_circ_0001772) (circRBM33) is implicated in the tumorigenesis of cancers. Nevertheless, the role of circRBM33 in CC is indistinct. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to evaluate the levels of circRBM33, miR-758-3p, and pumilio RNA binding family member 2 (PUM2) mRNA in tissue samples and cells. Cell proliferation, apoptosis, migration, invasion, and glycolysis were assessed using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, flow cytometry assay, transwell assay, or special commercial kits. Relative protein levels were examined via western blotting. The targeting relationship between circRBM33 or PUM2 and miR-758-3p was verified via dual-luciferase reporter or RNA pull-down assays. The role of circRBM33 was confirmed via tumor formation experiments. CircRPPH1 and PUM2 were upregulated while miR-758-3p was downregulated in CC tissues and cells. Functionally, circRBM33 knockdown constrained tumor growth in vivo and cured CC cell proliferation, migration, invasion, glycolysis, and fostered CC cell apoptosis in in vitro. Mechanistically, circRBM33 sponged miR-758-3p to modulate PUM2 expression. MiR-758-3p inhibitor neutralized circRBM33 silencing-mediated effects on the malignant behaviors of CC cells. PUM2 elevation overturned the suppressive influence of miR-758-3p upregulation on the malignant behaviors of CC cells. CircRBM33 fostered CC advancement via absorbing miR-758-3p and upregulating PUM2, indicating that circRBM33 was a possible target for CC treatment.

摘要

宫颈癌(CC)是一种妇科恶性肿瘤。环状 RNA(hsa_circ_0001772)(circRBM33)参与了癌症的发生。然而,circRBM33 在 CC 中的作用尚不清楚。采用实时定量聚合酶链反应(qRT-PCR)检测组织样本和细胞中 circRBM33、miR-758-3p 和 pumilio RNA 结合家族成员 2(PUM2)mRNA 的水平。采用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法、流式细胞术、transwell 法或特殊商业试剂盒评估细胞增殖、凋亡、迁移、侵袭和糖酵解。通过 Western blot 检测相对蛋白水平。通过双荧光素酶报告或 RNA 下拉实验验证 circRBM33 或 PUM2 与 miR-758-3p 的靶向关系。通过肿瘤形成实验证实 circRBM33 的作用。CC 组织和细胞中 circRPPH1 和 PUM2 上调,而 miR-758-3p 下调。功能上,circRBM33 敲低抑制体内肿瘤生长,体外治愈 CC 细胞增殖、迁移、侵袭、糖酵解,并促进 CC 细胞凋亡。机制上,circRBM33 吸附 miR-758-3p 调节 PUM2 表达。miR-758-3p 抑制剂中和了 circRBM33 沉默对 CC 细胞恶性行为的影响。PUM2 上调逆转了 miR-758-3p 上调对 CC 细胞恶性行为的抑制作用。circRBM33 通过吸附 miR-758-3p 上调 PUM2 促进 CC 进展,表明 circRBM33 可能是 CC 治疗的靶点。

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