Abstract: Objective To investigate the upstream transcription mechanism of acylglycerol kinase (AGK) in T lymphocyte leukemia, construct recombinant plasmids with different truncated fragments of the AGK gene promoter and detect their transcriptional activity. Methods Public databases such as TIMER2.0, The Human Protein Atlas, and Gene Expression Profiling Interactive Analysis were used to explore the role of AGK in non⁃solid tumors. Using the pGL4. 20⁃pAGK promoter luciferase full⁃length plasmid as a template, PCR was used to amplify truncated fragments of different lengths of the AGK promoter. The amplified fragments were inserted into the pGL4. 20⁃basic vector respectively to construct recombinant plasmids with different truncated fragments of the AGK promoter. After double enzyme digestion and sequence identification were correct, the recombinant plasmid was electroporated and transfected into human T lymphocytic leukemia cell Jurkat, and the dual⁃luciferase reporter gene assay was used to determine the dual luciferase activity of different truncated fragments of the AGK promoter. Results The expression of AGK was up⁃regulation in a variety of tumor types (P<0.05), and the expression level of AGK in leukemia cells was at a higher level. Log⁃rank test showed that the overall survival of patients with high AGK expression was lower than that of patients with low AGK expression (P=0.028）. Three high⁃scoring binding regions of transcription factor TEAD and AGK were predicted by JASPAR website, and the recombinant plasmids containing different truncated fragments of the AGK gene promoter were successfully constructed. Dual luciferase reporter gene assay showed that the transcriptional activity of AGK promoter was higher in the -540 bp to -80 bp region in T lymphocyte leukemia cells. Conclusions The transcription activity of the AGK gene promoter in the -540 bp to -80 bp region was higher in human T lymphocytic leukemia cells, laying the foundation for further research on the transcription and upstream regulatory mechanisms of AGK in T lymphocyte leukemia.

Key words: T lymphocyte leukemia, Acylglycerol kinase, Transcription factor, Recombinant plasmids, Luciferase reporter gene