Department of Anesthesiology, The First Affiliated Hospital of Nanchang University, Nanchang, 330006, Jiangxi, PR China.
Department of Anesthesiology, The First Affiliated Hospital of Nanchang University, Nanchang, 330006, Jiangxi, PR China.
Pathol Res Pract. 2020 Apr;216(4):152856. doi: 10.1016/j.prp.2020.152856. Epub 2020 Feb 11.
Our previous study states that propofol suppresses proliferation and migration of papillary thyroid cancer (PTC) cells by downregulation of lncRNA ANRIL. This study intended to probe the downstream mechanism of ANRIL in PTC with potential microRNAs (miR) and genes.
ANRIL expression was detected in normal thyroid epithelial cells (Nthy-ori 3-1) and PTC cells (TPC-1, FTC-133, K1 and BCPAP). ANRIL expression was inhibited in TPC-1 and BCPAP cells to explore the effects of si-ANRIL in PTC malignant behaviors. The gain-and loss-of functions of ANRIL/miR-320a were performed to measure their roles in PTC. Levels of ANRIL, miR-320a, HMGB1, apoptosis- and Wnt/β-catenin and NF-κB pathways-related proteins were measured. Dual-luciferase reporter gene assay and RNA pull-down assay were applied to verify ANRIL/miR-320a/HMGB1 relation. si-ANRIL was transplanted into xenograft tumors in nude mice.
ANRIL was upregulated in TPC-1 and BCPAP cells. miR-320a targeted HMGB1, and ANRIL bound to miR-320a. In TPC-1 and BCPAP cells, si-ANRIL prevented PTC cell malignant behaviors, and inactivated the Wnt/β-catenin and NF-κB pathways; while si-ANRIL + miR-320a inhibition showed opposite trends. Overexpressing miR-320a promoted malignant behaviors of TPC-1 cells. In 6 μg/mL propofol-treated TPC-1 cells, miR-320a inhibition weakened propofol's inhibitory effects on PTC cell growth. After ANRIL inhibition, the volume and weight of xenograft tumors were decreased.
Propofol upregulated miR-320a and reduced HMGB1 by downregulating ANRIL and inactivating the Wnt/β-catenin and NF-κB pathways, thus preventing PTC cell malignant behaviors. This study may offer new insights in PTC prevention and treatment.
我们之前的研究表明，丙泊酚通过下调长链非编码 RNA ANRIL 抑制甲状腺乳头状癌（PTC）细胞的增殖和迁移。本研究旨在探讨 ANRIL 在 PTC 中的潜在微小 RNA（miR）和基因下游机制。
检测正常甲状腺上皮细胞（Nthy-ori 3-1）和 PTC 细胞（TPC-1、FTC-133、K1 和 BCPAP）中的 ANRIL 表达。抑制 TPC-1 和 BCPAP 细胞中的 si-ANRIL，以探讨 si-ANRIL 在 PTC 恶性行为中的作用。进行 ANRIL/miR-320a 的增益和失活功能，以测量它们在 PTC 中的作用。测量 ANRIL、miR-320a、HMGB1、凋亡以及 Wnt/β-catenin 和 NF-κB 通路相关蛋白的水平。应用双荧光素酶报告基因检测和 RNA 下拉实验验证 ANRIL/miR-320a/HMGB1 关系。将 si-ANRIL 移植到裸鼠的异种移植肿瘤中。
TPC-1 和 BCPAP 细胞中 ANRIL 上调。miR-320a 靶向 HMGB1，ANRIL 与 miR-320a 结合。在 TPC-1 和 BCPAP 细胞中，si-ANRIL 可防止 PTC 细胞的恶性行为，并使 Wnt/β-catenin 和 NF-κB 通路失活；而 si-ANRIL+miR-320a 抑制则表现出相反的趋势。过表达 miR-320a 促进了 TPC-1 细胞的恶性行为。在 6μg/ml 丙泊酚处理的 TPC-1 细胞中，miR-320a 抑制削弱了丙泊酚对 PTC 细胞生长的抑制作用。抑制 ANRIL 后，异种移植瘤的体积和重量均减少。
丙泊酚通过下调 ANRIL 并使 Wnt/β-catenin 和 NF-κB 通路失活，上调 miR-320a 并降低 HMGB1，从而防止 PTC 细胞的恶性行为。本研究为 PTC 的预防和治疗提供了新的思路。
