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Archive > Volume 31 Issue 9 > 2024,31(9):857-863. DOI:10.3872/j.issn.1007-385X.2024.09.003 Prev Next
Basic Research
Effects of MET knockdown on proliferation, migration and sensitivity to 5-FU and cisplatin of laryngeal cancer Hep-2 cells
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Abstract:
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.
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