Department of Obstetrics and Gynecology, The First Affiliated Hospital of Soochow University, 188 Shizi Road, Suzhou, 215006, Jiangsu, People's Republic of China.
Department of Obstetrics and Gynecology, The First People's Hospital of Yancheng, Yancheng, 224001, Jiangsu, People's Republic of China.
Biol Res. 2020 Mar 10;53(1):10. doi: 10.1186/s40659-020-00277-4.
The aim of this study was to investigate the effect role and mechanism of miR-30b-3p on ovarian cancer cells biological function.
The expression of miR-30b-3p was detected in ovarian cancer cell lines and normal ovarian epithelial cell line by qRT-PCR. Mir-30b-3p mimic was transfected into OVCAR3 cells. Cell-counting kit-8 (CCK-8) assay was conducted to explore the effect of mir-30b-3p on the OVCAR3 cells' proliferation. Cell cycle and apoptosis were detected by Flow cytometry. Cell invasion ability was detected by Transwell test. The regulation of putative target of miR-30b-3p was verified by double luciferase reporter assays and Western blot.
We found that miR-30b-3p was downregulated in OVCAR3 cells. Overexpression of miR-30b-3p suppressed proliferation, promoted apoptosis, slowed cell cycle and inhibited migration and invasion of OVCAR3 cells. Bioinformatics analysis identified 3'-untranslated region (3'UTR) of Collagen triple helix repeat-containing 1 (CTHRC1) as the presumed binding site for miR-30b-3p. Detection of double luciferase reporter and Western-Blot result confirmed that CTHRC1 was the target gene of miR-30b-3p. Furthermore, E-cadherin, β-cadherin and Vimentin protein expression level were changed after transfection of miR-30b-3p.
miR-30b-3p function as an anti-cancer gene. Overexpression of miR-30b-3p can inhibit the biological function of ovarian cancer cells. MiR-30b-3p targets CTHRC1 gene plays an important role in epithelial-mesenchymal transformation (EMT), and supports miR-30b-3p as a potential biological indicator for ovarian cancer in the future.
本研究旨在探讨 miR-30b-3p 在卵巢癌细胞生物学功能中的作用和机制。
通过 qRT-PCR 检测卵巢癌细胞系和正常卵巢上皮细胞系中 miR-30b-3p 的表达。将 miR-30b-3p 模拟物转染至 OVCAR3 细胞中。通过细胞计数试剂盒-8（CCK-8）测定法研究 miR-30b-3p 对 OVCAR3 细胞增殖的影响。通过流式细胞术检测细胞周期和凋亡。通过 Transwell 试验检测细胞侵袭能力。通过双荧光素酶报告基因检测和 Western blot 验证潜在靶基因的调控。
我们发现 miR-30b-3p 在 OVCAR3 细胞中下调。miR-30b-3p 的过表达抑制增殖，促进凋亡，减缓细胞周期并抑制 OVCAR3 细胞的迁移和侵袭。生物信息学分析鉴定出 Collagen triple helix repeat-containing 1（CTHRC1）的 3'-非翻译区（3'UTR）为 miR-30b-3p 的假定结合位点。双荧光素酶报告基因和 Western blot 检测结果证实 CTHRC1 是 miR-30b-3p 的靶基因。此外，转染 miR-30b-3p 后 E-钙黏蛋白、β-钙黏蛋白和波形蛋白的蛋白表达水平发生改变。
miR-30b-3p 作为一种抗癌基因发挥作用。miR-30b-3p 的过表达可抑制卵巢癌细胞的生物学功能。miR-30b-3p 靶向 CTHRC1 基因在上皮间质转化（EMT）中发挥重要作用，支持 miR-30b-3p 作为未来卵巢癌的潜在生物学标志物。
