Department of Oncology, Taizhou Clinical Medical School of Nanjing Medical University, Taizhou, Jiangsu Province, 225300, China; Department of Oncology, The People's Hospital of Taizhou, Taizhou, Jiangsu Province, 225300, China.
Institute of Clinical Medicine, Taizhou Clinical Medical School of Nanjing Medical University, Taizhou, Jiangsu Province, 225300, China.
Pathol Res Pract. 2021 Jan;217:153289. doi: 10.1016/j.prp.2020.153289. Epub 2020 Nov 15.
Long non-coding RNA (lncRNA) Growth-Arrest Associated LncRNA 1 (GASL1) is a lncRNA with a suppressive role in glioma, prostate carcinoma and gastric carcinoma, whereas its involvement in esophageal cancer is unknown. In the present study, we used RT-qPCR to detect the expression of GASL1 in esophageal cancer cell carcinoma (ESCC) cell lines, and constructed the overexpression and interference plasmids of GASL1 and the interference plasmid of DKK1. CCK8 was used to detect the cell proliferation level, clone formation experiment was used to detect the cell clonal formation ability, flow cytometry was used to detect the cell cycle level, and wound healing and Transwell experiments were respectively used to detect the cell invasion and migration. The interference and overexpression plasmids of GASL1 were injected into mice subcutaneously for tumor-bearing experiment. The body weight, tumor growth curve, and tumor weight of mice were recorded, and western blot was used to detect the expression of proliferation-, invasion-, and migration-related proteins and the expression of Wnt3a/β-catenin signal-related proteins in tumor tissues. LncRNA GASL1 was down-regulated in ESCC cell lines, and GASL1 inhibited ESCC cell progression and regulated cell cycle arrest in ESCC cells. In vivo, GASL1 inhibited tumor growth. GASL1 decreased the protein levels of DDK1, Wnt3a, β-catenin, and c-MYC in ESCC cell lines. Interfering DKK1 activates Wnt3a/β--catenin signal to reverse the inhibitory effects of GASL1 on proliferation, cell cycle acceleration, invasion, and migration. In conclusion, lncRNA GASL1 regulates cell migration, invasion and cell cycle stagnation by inactivating the wnt/β-catenin signaling.
长链非编码 RNA（lncRNA）生长停滞相关长链非编码 RNA 1（GASL1）在神经胶质瘤、前列腺癌和胃癌中起抑制作用，但其在食管癌中的作用尚不清楚。在本研究中，我们使用 RT-qPCR 检测了食管癌细胞系（ESCC）中 GASL1 的表达，并构建了 GASL1 的过表达和干扰质粒以及 DKK1 的干扰质粒。CCK8 用于检测细胞增殖水平，克隆形成实验用于检测细胞克隆形成能力，流式细胞术用于检测细胞周期水平，划痕愈合和 Transwell 实验分别用于检测细胞侵袭和迁移。将 GASL1 的干扰和过表达质粒注入小鼠皮下进行荷瘤实验。记录小鼠的体重、肿瘤生长曲线和肿瘤重量，并使用 Western blot 检测肿瘤组织中增殖、侵袭和迁移相关蛋白的表达以及 Wnt3a/β-catenin 信号相关蛋白的表达。lncRNA GASL1 在 ESCC 细胞系中下调，GASL1 抑制 ESCC 细胞进展并调节 ESCC 细胞周期停滞。在体内，GASL1 抑制肿瘤生长。GASL1 降低了 ESCC 细胞系中 DDK1、Wnt3a、β-catenin 和 c-MYC 的蛋白水平。干扰 DKK1 激活 Wnt3a/β-catenin 信号，逆转 GASL1 对增殖、细胞周期加速、侵袭和迁移的抑制作用。总之，lncRNA GASL1 通过失活 wnt/β-catenin 信号调节细胞迁移、侵袭和细胞周期停滞。
