Screening of human membrane protein genes activating MMPs highly expressed in squamous-cell carcinoma of head and neck
XU Hongbo
MI Guangxi
HUANG Junwei
LIU Zeyang
YIN Gaofei
HUANG Zhigang
Abstract：Objective To screen out the human membrane protein genes activating matrix metalloproteinases (MMPs) (MMP1, MMP2, MMP3, MMP9, MMP10, MMP12, and MMP13) highly expressed in squamous-cell carcinoma of head and neck.Methods Seven alternative MMP promoter reporter gene plasmids were constructed.PMA, a kind of protein kinase C activator, was used to stimulate them;Dual luciferase assay system was used to detect the basic and stimulated activities of MMP promoter reporters.The relative luciferase activity (RLA) values were measured as the ratio of the basic divided by the stimulated ones;the highest RLA was selected MMP promoter reporter gene plasmid.A total of 722 candidate genes encoding potential human transmembrane proteins were screened in a dual luciferase based assay system.The candidate genes with RLA greater than 10 were considered as the activator to MMP promoter reporters in the first round, and the second and third rounds followed the same way.Finally, the efficacious human membrane protein genes were ascertained.Results The RLAs of MMP1, MMP2, MMP3, MMP9, MMP10, MMP12, and MMP13 promoter reporters stimulated by PMA were 4.53±0.72, 1.47±0.28, 23.28±1.96, 2.37±0.46, 8.81±1.35, 2.15±0.51, and 3.47±0.99.The MMP3 reporter was selected for a high through-put screening for the sensitivity (P<0.05).G protein-coupled receptor 65 (GPR65), AXL receptor tyrosine kinase (AXL), and tumor necrosis factor receptor superfamily 10b (TNFRSF10B) were confirmed as significant activators to MMPs by screening all 722 plasmids in 3 rounds.Conclusion GPR65, AXL, and TNFRSF10B could activate MMPs highly expressed in head and neck squamous-cell carcinoma.
Keywords：head and neck squamous-cell carcinomamatrix metalloproteinaseshuman membrane proteinG protein-coupled receptor 65AXL receptor tyrosine kinasetumor necrosis factor receptor superfamily 10b
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 1-3 )