Wang Yue, Guo Yubing, Lu Yanyuan, Sun Yeqing, Xu Dan
Institute of Environmental Systems Biology, Environment Science and Engineering College, Dalian Maritime University, Linghai Road 1, Dalian, 116026, PR China.
Institute of Environmental Systems Biology, Environment Science and Engineering College, Dalian Maritime University, Linghai Road 1, Dalian, 116026, PR China.
Sci Total Environ. 2022 Nov 1;845:157252. doi: 10.1016/j.scitotenv.2022.157252. Epub 2022 Jul 8.
Endosulfan belongs to persistent organic pollutants (POPs), closely related to an increased risk of prostate cancer (PCa). The existing evidence shows that lncRNAs compete with miRNAs for binding sites and contribute to the onset and progression of human malignancies. In this study we investigate how endosulfan promotes cell migration and invasion in DU145 and PC3 prostate cancer cells through epigenetic mechanism of lncRNA-miRNA regulation. Based on our past research we focused on PTP4A3 and constructed wild-type (WT) and mutant PTP4A3 plasmids for further analysis. Our results revealed that transfection of PTP4A3-WT can lead to changes in the expression of epithelial-mesenchymal transition (EMT) biomarkers and critical proteins in the TGF-β signaling pathway, and promote cell migration and invasion in PCa cells. Bioinformatics analysis shows that there were complementary sequences in PTP4A3 3'-UTR and KCNQ1OT1 3'-UTR to the seed sequence of hsa-miR-137-3p, and dual luciferase reporter assay indicates the potential binding capacity of miR-137-3p to 3'-UTR of PTP4A3 and KCNQ1OT1. We found that miR-137-3p mimic inhibited cell migration and invasion, as well as repressed alterations of EMT biomarkers and critical proteins in the TGF-β signaling pathway. Rescue experiment results revealed that co-transfection of miR-137-3p mimic and PTP4A3-WT plasmid reversed these changes following transfection with miR-137-3p mimic alone. We found that KCNQ1OT1 was predominantly distributed in the cytoplasm from a subcellular fractionation assay. Functionally, silencing of KCNQ1OT1 repressed cell migration and invasion, and caused alterations of EMT biomarkers and critical proteins in the TGF-β signaling pathway, which were all restored by co-transfection with anti-miR-137-3p or PTP4A3-WT plasmid. Furthermore, overexpression of miR-137-3p or silencing of KCNQ1OT1 dramatically rescued the effects of endosulfan on promoting cell migration and invasion. These findings suggest that endosulfan can indeed promote cell migration and invasion via the KCNQ1OT1/miR-137-3p/PTP4A3 axis in PCa cells.
硫丹属于持久性有机污染物（POPs），与前列腺癌（PCa）风险增加密切相关。现有证据表明，长链非编码RNA（lncRNAs）与微小RNA（miRNAs）竞争结合位点，并参与人类恶性肿瘤的发生和发展。在本研究中，我们探究硫丹如何通过lncRNA-miRNA调控的表观遗传机制促进DU145和PC3前列腺癌细胞的迁移和侵袭。基于我们过去的研究，我们聚焦于PTP4A3，并构建了野生型（WT）和突变型PTP4A3质粒用于进一步分析。我们的结果显示，转染PTP4A3-WT可导致上皮-间质转化（EMT）生物标志物和TGF-β信号通路关键蛋白的表达发生变化，并促进PCa细胞的迁移和侵袭。生物信息学分析表明，PTP4A3 3'-UTR和KCNQ1OT1 3'-UTR中存在与hsa-miR-137-3p种子序列互补的序列，双荧光素酶报告基因检测表明miR-137-3p与PTP4A3和KCNQ1OT1的3'-UTR具有潜在结合能力。我们发现，miR-137-3p模拟物抑制细胞迁移和侵袭，以及抑制EMT生物标志物和TGF-β信号通路关键蛋白的改变。挽救实验结果显示，共转染miR-137-3p模拟物和PTP4A3-WT质粒可逆转单独转染miR-137-3p模拟物后的这些变化。我们通过亚细胞分级分离实验发现KCNQ1OT1主要分布在细胞质中。在功能上，沉默KCNQ1OT1可抑制细胞迁移和侵袭，并导致EMT生物标志物和TGF-β信号通路关键蛋白的改变，而共转染抗miR-137-3p或PTP4A3-WT质粒可恢复这些改变。此外，miR-137-3p的过表达或KCNQ1OT1的沉默显著挽救了硫丹对促进细胞迁移和侵袭的作用。这些发现表明，硫丹确实可通过PCa细胞中的KCNQ1OT1/miR-137-3p/PTP4A3轴促进细胞迁移和侵袭。
