Department of Medical Oncology, Dalian Municipal Central Hospital Affiliated of Dalian Medical University, Dalian 116033, China.
Math Biosci Eng. 2019 May 23;16(5):4607-4621. doi: 10.3934/mbe.2019231.
To explore the potential mechanism which miR-527 targeting the heparan sulfate 6-O-endosulfatase (SULF2) regulates TGF-β/SMAD signaling pathway induced epithelial-mesenchymal transition (EMT) in non-small-cell lung cancer (NSCLC).
38 pairs of lung tumor biopsies and corresponding paracancerous biopsies were obtained from NSCLC patients with surgical resection, normal human bronchial epithelial BEAS-2B cells and five NSCLS cell lines were applied for our study. miR-527 and SULF2 expression were determined by qRT-PCR and immunohistochemistry. MiR-527 and SULF2 biological link were predicted by Targetscan.org and tested by dual luciferase. Cells proliferation and apoptosis were respectively detected by EDU staining and flow cytometry. Cells migration was examined by transwell and scratch-wound assay. Expression of proteins related to EMT and TGF-β/SMAD signaling pathway, such as E-cadherin, N-cadherin, p-Samd3 and p-Smad2, was detected by western blot.
miR-527 expression was decreased in lung tumor tissues and NSCLS cell lines, conversely, SULF2 expression was significantly increased. In addition, we found that miR-527 targeted 3'-untranslated regions (3'-UTR) of SULF2 and mediated its expression. Overexpression of miR-527 evidently suppressed NSCLC proliferation, invasion and EMT via TGF-β/SMAD signaling pathway. Moreover, the silence of SULF2 exhibited a similar effect.
miR-527 targeting SULF2 down-regulated SULF2 expression, concurrently, suppressed NSCLC epithelial-mesenchymal transition and invasion via inhibiting TGF-β/SMAD signaling pathway.
探讨 miR-527 靶向硫酸乙酰肝素 6-O-硫酸酯酶（SULF2）调控 TGF-β/SMAD 信号通路诱导非小细胞肺癌（NSCLC）上皮-间充质转化（EMT）的潜在机制。
收集 38 对 NSCLC 患者手术切除的肿瘤组织及其癌旁组织，正常人类支气管上皮 BEAS-2B 细胞和 5 种 NSCLC 细胞系用于本研究。通过 qRT-PCR 和免疫组化检测 miR-527 和 SULF2 的表达。Targetscan.org 预测 miR-527 和 SULF2 的生物学联系，并通过双荧光素酶实验进行验证。通过 EDU 染色和流式细胞术分别检测细胞增殖和凋亡。通过 Transwell 和划痕实验检测细胞迁移。通过 Western blot 检测 EMT 和 TGF-β/SMAD 信号通路相关蛋白（如 E-cadherin、N-cadherin、p-Samd3 和 p-Smad2）的表达。
miR-527 在肺癌组织和 NSCLC 细胞系中的表达降低，而 SULF2 的表达明显升高。此外，我们发现 miR-527 靶向 SULF2 的 3'-非翻译区（3'-UTR）并调节其表达。miR-527 的过表达通过 TGF-β/SMAD 信号通路显著抑制 NSCLC 的增殖、侵袭和 EMT。此外，沉默 SULF2 也表现出类似的效果。
miR-527 靶向 SULF2 下调 SULF2 的表达，通过抑制 TGF-β/SMAD 信号通路，同时抑制 NSCLC 的上皮-间充质转化和侵袭。
