Experimental study on the effects of m7G methyltransferase 1 regulating lymphocyte activator gene-3 on the proliferation,migration and invasion of head and neck squamous cell carcinoma cells
WANG Jingxiu
ZHOU Jingjie
LI Jianru
LYU Shujun
Abstract：Objective To investigate the effects and possible mechanisms of m7G methyltransferase 1(METTL1)regulating the expression of lymphocyte activator gene-3(LAG-3)on the biological behaviors such as proliferation,migration and invasion of head and neck squamous cell carcinoma(HNSCC)cells.Methods Fifty cases each of HNSCC tissues surgically resected in Jingmen Central Hospital from May 2022 to May 2024 and the corresponding adjacent normal tissues were collected.FaDu cells of HNSCC were divided into different treatment groups:siRNA negative control(si-NC group),METTL1 knockdown(si-METTL1 group),co-transfection of si-METTL1 and over-expression negative control(si-METTL1+oe-NC group),and co-transfection of si-METTL1 and LAG-3 over-expression(si-METTL1+oe-LAG-3 group).Western blotting was used to detect the expression of METTL1 and LAG-3 in tissues and cells.Experimental analysis of the targeting relationship between METTL1 and LAG-3 using dual luciferase reporter genes.The CCK-8 method and EdU method were used to detect the proliferation ability of each group of cells,and the Transwell chamber experiment was used to detect the migration and invasion ability of each group of cells.Western blotting was used to detect the expression of proliferating cell nuclear antigen(PCNA)and epithelial mesenchymal transition(EMT)related proteins(E-cadherin,N-cadherin)in each group of cells.Results Compared with HOK cells and adjacent normal tissues,the protein expression level of METTL1 was significantly higher in FaDu cells and HNSCC tissues[(1.00±0.08)vs.(1.58±0.11),(1.00±0.10)vs.(2.34±0.12)],and the protein expression level of LAG-3 was also significantly higher[(1.02±0.10)vs.(1.94±0.13),(1.01±0.09)vs.(1.87±0.11)].All the above differences were statistically significant(P<0.05).The dual-luciferase assay confirmed a targeted regulatory relationship between METTL1 and LAG-3.Compared with the si-NC group,the si-METTL1 group showed a significant decrease in METTL1 protein expression[(1.00±0.10)vs.(0.39±0.08)]and a significant increase in LAG-3 protein expression[(1.00±0.09)vs.(1.84±0.13)].Meanwhile,in the si-METTL1 group,the cell proliferation rate[(53.56±6.07)%vs.(22.67±4.25)%],number of migrated cells[(80.35±9.10)vs.(33.84±5.27)],number of invaded cells[(89.48±11.05)vs.(41.56±4.27)],PCNA protein[(1.00±0.11)vs.(0.61±0.08)]and N-cadherin protein[(1.00±0.10)vs.(0.53±0.08)]were significantly reduced(all P<0.05),while the protein expression level of E-cadherin[(1.00±0.10)vs.(1.37±0.12)]was significantly increased(P<0.05).Compared with the si-METTL1+oe-NC group,the si-METTL1+oe-LAG-3 group exhibited significantly higher LAG-3 protein expression[(1.77±0.11)vs.(2.94±0.13)],cell proliferation rate[(21.57±4.31)%vs.(51.28±6.57)%],number of migrated cells[(32.16±5.65)vs.(69.15±6.02)],number of invaded cells[(43.08±4.09)vs.(73.68±8.91)],the expression of PCNA protein[(0.59±0.07)vs.(0.95±0.10)]and N-cadherin protein[(0.55±0.10)vs.(0.89±0.09)](all P<0.05),whereas the protein expression level of E-cadherin[(1.40±0.10)vs.(0.98±0.09)]was significantly decreased(P<0.05).Conclusion METTL1 is highly expressed in HNSCC and regulates the proliferation,migration,and invasion of HNSCC cells by targeting LAG-3.
Keywords：Head and neck squamous cell carcinomam7G methyltransferase 1Lymphocyte activating gene-3Cell biology
Publication Date：2025-08-28
Online Publishing Date：2025-10-21(First online date of this platform, not the publication date of the document)
Pages：6( 735-740 )