目的：探究敲减间质表皮转化因子（MET）表达对人喉鳞状细胞癌（LSCC）Hep-2细胞生长、迁移及5-氟尿嘧啶（5-FU） 和顺铂敏感性的影响。方法：通过基因表达综合数据库（GEO）及癌症基因组图谱（TCGA）数据库数据分析人 LSCC 组织中 MET mRNA的表达水平；常规培养正常人支气管上皮细胞16HBE与人LSCC细胞Hep-2、KBV200和TU212，采用qPCR法和WB 法检测16HBE、Hep-2、KBV200和TU212细胞中MET基因和蛋白的表达水平。用LipofectamineTM 3000将MET敲减质粒（si-Met） 和对照质粒（si-NC）转染至Hep-2细胞中，分为空白对照组，si-NC组和si-Met组。采用MTT法、流式细胞术、划痕愈合实验分别 检测各组Hep-2细胞的增殖、迁移能力、周期分布以及对5-FU和顺铂的敏感性。结果：数据库数据分析显示LSCC组织中MET mRNA呈高表达（P < 0.05），Hep-2、KBV200和TU212 细胞中 MET mRNA 和蛋白的表达水平也均明显高于 16HBE 细胞（均 P < 0.01）。敲减MET表达后，Hep-2细胞中MET mRNA及蛋白表达水平均明显降低（P < 0.01或P < 0.001）、细胞增殖活力显著 下降（P < 0.000 1）、G0/G1期细胞数量明显升高（P < 0.000 1）、S期细胞数量明显降低（P < 0.000 1）。敲减MET表达后，不同浓度 5-FU或顺铂对细胞增殖的抑制率均显著升高、药物半数抑制浓度（IC50 ）均降低（均 P < 0.000 1），划痕愈合率明显降低、迁移能 力下降（均P < 0.05）。结论：MET在人LSCC组织和细胞中呈高表达，敲减MET可有效抑制Hep-2细胞中MET的表达，抑制细 胞增殖、迁移能力，使其周期阻滞于G1期，增强Hep-2细胞对5-FU和顺铂的敏感性。

Objective: To investigate the effects of mesenchymal to epithelial transition factors (MET) knockdown on the proliferation, migration, and sensitivity to 5-FU and cisplatin in human laryngeal squamous cell carcinoma (LSCC) Hep-2 cells. Methods: The expression of MET mRNA in LSCC tissues was analyzed using data from the Gene Expression Omnibus (GEO) and The Cancer and Tumor Gene Atlas (TCGA). Human normal bronchial epithelial cells (16HBE) and human LSCC cells (Hep-2, KBV200 and TU212) were routinely cultured, and the expression levels of MET in these cells were detected by qPCR and WB assay. The MET knockout plasmid (si-Met) and control plasmid (si-NC) were transfected into Hep-2 cells using LipofectamineTM 3000, and the cells were divided into blank control group, si-NC group and si-Met group. The proliferation, migration, cell cycle distribution, and sensitivity to 5-FU and cisplatin of Hep-2 cells in each group were detected by MTT assay, flow cytometry and scratch assay, respectively. Results: Database analysis showed high expression of MET mRNA in LSCC tissue (P < 0.05). The mRNA and protein expression levels of MET in Hep2 cells, KBV200 cells and TU212 cells were significantly higher than those in 16HBE cells (all P < 0.01). After MET knockdown, the mRNA and protein levels of MET in Hep-2 cells were significantly reduced (P < 0.01 or P < 0.001), cell proliferation activity was significantly decreased (P < 0.000 1), the number of G0/G1 phase cells was significantly increased (P < 0.000 1), and the number of S phase cells was significantly reduced. Additionally, after MET knockdown, the inhibitory rates of cell proliferation by different concentrations of 5-FU or cisplatin were significantly enhanced, and the half maximal inhibitory concentration (IC50) was reduced (all P < 0.000 1). The scratch healing rate and migration ability were significantly reduced (all P < 0.05). Conclusion: MET is highly expressed in human LSCC tissues and cells. Knocking down MET can effectively inhibit the expression of MET in Hep-2 cells, suppress cell proliferation and migration ability, arrest the cell cycle in G1 phase, and enhance the sensitivity of Hep-2 cells to 5-FU and cisplatin.

河北省卫生厅重点科技研究计划课题（No.20220237）