First Department of Lung Cancer Chemotherapy, Affiliated Cancer Hospital of Xinjiang Medical University, Urumqi 830011, PR China; Department of Hematology and Oncology, Shenzhen University General Hospital, Shenzhen 518055, PR China.
Department of Gastrointestinal Surgery, Affiliated Cancer Hospital of Xinjiang Medical University, Urumqi 830011, PR China.
Biomed Pharmacother. 2019 Mar;111:476-484. doi: 10.1016/j.biopha.2018.12.057. Epub 2018 Dec 27.
Esophageal squamous cell carcinoma (ESCC) is an aggressive tumor entity characterized by early metastasis and late diagnosis. MicroRNA-133b (miR-133b) has been considered as a tumor suppressor in many human cancers by regulating epidermal growth factor receptor (EGFR). However, the specific effects of miR-133b and EGFR on ESCC remain unclear.
qRT-PCR and western blotting were applied for measuring expression of mRNA and protein. Flow cytometry was used for detecting cell cycle and apoptosis. Cell proliferation, migration and invasion were detected by colony formation and transwell assays. Luciferase reporter assay was used to confirm the interaction between miR-133b and EGFR.
Low expression of miR-133b and high expression of EGFR were identified in ESCC cells and tissues. Overexpression of miR-133b or knockdown of EGFR suppressed the cell proliferation, migration, and invasion of ESCC cells, and raised the percentage of G1 phase cells. The apoptosis of ESCC cells were promoted by increasing miR-133b and decreasing EGFR expression. Luciferase reporter assay confirmed EGFR as the target of miR-133b in ESCC cells. Overexpression of miR-133b significantly decreased the phosphorylation of PI3K, ERK and AKT by directly down-regulating EGFR. Higher expression of E-cadherin and CK-18 and lower expression of Vimentin and N-cadherin were observed after the transfection of miR-133b mimics or shEGFR.
Overexpression of miR-133b could suppress proliferation, migration and invasion of ESCC cells by inhibiting MAPK/ERK and PI3K/AKT signaling pathways through targeting EGFR, indicating that miR-133b might be a potential therapeutic target for the treatment of ESCC.
食管鳞状细胞癌（ESCC）是一种侵袭性肿瘤实体，其特征在于早期转移和晚期诊断。MicroRNA-133b（miR-133b）通过调节表皮生长因子受体（EGFR）被认为是许多人类癌症的肿瘤抑制因子。然而，miR-133b 和 EGFR 对 ESCC 的具体影响仍不清楚。
qRT-PCR 和 Western blot 用于测量 mRNA 和蛋白的表达。流式细胞术用于检测细胞周期和细胞凋亡。通过集落形成和 Transwell 测定检测细胞增殖、迁移和侵袭。荧光素酶报告基因实验用于证实 miR-133b 和 EGFR 之间的相互作用。
ESCC 细胞和组织中发现 miR-133b 表达低，EGFR 表达高。过表达 miR-133b 或敲低 EGFR 抑制 ESCC 细胞的增殖、迁移和侵袭，并提高 G1 期细胞的比例。增加 miR-133b 和降低 EGFR 表达可促进 ESCC 细胞的凋亡。荧光素酶报告基因实验证实 EGFR 是 ESCC 细胞中 miR-133b 的靶标。过表达 miR-133b 可通过直接下调 EGFR 显著降低 PI3K、ERK 和 AKT 的磷酸化。转染 miR-133b 模拟物或 shEGFR 后，观察到 E-钙黏蛋白和 CK-18 的表达升高，波形蛋白和 N-钙黏蛋白的表达降低。
过表达 miR-133b 通过靶向 EGFR 抑制 MAPK/ERK 和 PI3K/AKT 信号通路，可抑制 ESCC 细胞的增殖、迁移和侵袭，表明 miR-133b 可能是 ESCC 治疗的潜在治疗靶点。
