Zhang Zhuo, Li Jingxia, Yan Bo, Tu Huailu, Huang Chao, Costa Max
Department of Environmental Medicine, New York University Grossman School of Medicine, 341 East 25 Street, New York, New York, 10010, USA.
Heliyon. 2022 Aug 12;8(8):e10086. doi: 10.1016/j.heliyon.2022.e10086. eCollection 2022 Aug.
Chronic exposure of human bronchial epithelial BEAS-2B cells to hexavalent chromium (Cr(VI)) causes malignant cell transformation. These transformed cells exhibit increases in migration and invasion. Neuronal precursor of developmentally downregulated protein 9 (NEDD9) is upregulated in Cr(VI)-transformed cells compared to that of passage-matched normal BEAS-2B cells. Knockdown of NEDD9 by its shRNA reduced invasion and migration of Cr(VI)-transformed cells. Maternally expressed gene 3 (MEG3), a long noncoding RNA, was lost and microRNA 145 (miR-145) was upregulated in Cr(VI)-transformed cells. MEG3 was bound to miR-145 and this binding reduced its activity. Overexpression of MEG3 or inhibition of miR-145 decreased invasion and migration of Cr(VI)-transformed cells. Overexpression of MEG3 was able to decrease miR-145 level and NEDD9 protein level in Cr(VI)-transformed cells. Ectopic expression of MEG3 was also shown to reduce β-catenin activation. Inhibition of miR-145 in Cr(VI)-transformed cells decreased Slug, an important transcription factor that regulates epithelial-to-mesenchymal transition (EMT). Inhibition of miR-145 was found to increase MEG3 in Cr(VI)-transformed cells. Further studies showed that mutation of MEG3 at the binding site for miR-145 did not change NEDD9 and failed to decrease invasion and migration. The present study demonstrated that loss of MEG3 and upregulation of miR-145 elevated NEDD9, resulting in activation of β-catenin and further upregulation of EMT, leading to increased invasion and migration of Cr(VI)-transformed cells.
人支气管上皮BEAS-2B细胞长期暴露于六价铬(Cr(VI))会导致恶性细胞转化。这些转化细胞的迁移和侵袭能力增强。与传代匹配的正常BEAS-2B细胞相比,发育下调蛋白9(NEDD9)的神经元前体在Cr(VI)转化细胞中上调。通过其短发夹RNA(shRNA)敲低NEDD9可降低Cr(VI)转化细胞的侵袭和迁移能力。母系表达基因3(MEG3)是一种长链非编码RNA,在Cr(VI)转化细胞中缺失,而微小RNA 145(miR-145)上调。MEG3与miR-145结合,这种结合降低了miR-145的活性。MEG3的过表达或miR-145的抑制可降低Cr(VI)转化细胞的侵袭和迁移能力。MEG3的过表达能够降低Cr(VI)转化细胞中miR-145水平和NEDD9蛋白水平。MEG3的异位表达还显示可降低β-连环蛋白的激活。在Cr(VI)转化细胞中抑制miR-145可降低Slug,Slug是一种调节上皮-间质转化(EMT)的重要转录因子。发现在Cr(VI)转化细胞中抑制miR-145可增加MEG3。进一步研究表明,MEG3在miR-145结合位点的突变不会改变NEDD9,也无法降低侵袭和迁移能力。本研究表明,MEG3的缺失和miR-145的上调升高了NEDD9,导致β-连环蛋白激活并进一步上调EMT,从而导致Cr(VI)转化细胞的侵袭和迁移增加。