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Mir-30b-3p affects the migration and invasion function of ovarian cancer cells by targeting the CTHRC1 gene

机译:miR-30b-3p通过靶向CTHRC1基因来影响卵巢癌细胞的迁移和侵袭功能

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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.
机译:本研究的目的是探讨miR-30b-3p对卵巢癌细胞生物功能的影响作用和机制。通过QRT-PCR在卵巢癌细胞系和正常卵巢上皮细胞系中检测miR-30b-3p的表达。将miR-30b-3p模拟物转染到ovcar3细胞中。进行细胞计数试剂盒-8(CCK-8)测定以探讨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细胞凋亡。通过Transwell测试检测细胞侵入能力。通过双荧光素酶报告结果和Western印迹验证了MiR-30B-3P的推定靶标的调节。我们发现miR-30b-3p在ovcar3细胞中下调。 MIR-30B-3P的过度表达抑制了增殖,促进细胞凋亡,减缓细胞周期,抑制了OVCAR3细胞的迁移和侵袭。生物信息学分析确定了含有胶原三重螺旋重复的1(CTHRC1)的3'-未旋转的区域(3'UTR)作为MIR-30B-3P的假定结合位点。检测双荧光素酶报告和蛋白质印迹结果证实,CTHRC1是miR-30b-3p的靶基因。此外,在转染miR-30b-3p后改变e-cadherin,β-cadherin和平节蛋白表达水平。 miR-30b-3p用作抗癌基因。 miR-30b-3p的过度表达可以抑制卵巢癌细胞的生物学功能。 MiR-30B-3P靶标CTHRC1基因在上皮 - 间充质转化(EMT)中起重要作用,并支持未来卵巢癌的潜在生物学指标的miR-30b-3p。

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