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The Chinese Journal of Dental Research

Year 2004
Volume 7 , Issue 2

Pages: 37 - 40

An Apotosis-Mediation Study in Tongue Carcinoma Cells with a Combination of Tumor Necrosis Factor Receptor-1 (TNFR-1) Gene Transfection and Recombinant Human TNF-alpha

Da Qing Liu, DDS, PhD/Shu Xia Zhou, DDS/Zhen Qiang Situ, DDS/Cheng Ji Wang, PhD/An Gang Yang, PhD

Objective: To construct the cell line T-T expressing the activity of tumor necrosis factor receptor-1 (TNFR-1) protein, and to analyze the effect and mechanism in which T-T cells treated with recombinant human tumor necrosis factor-alpha (rhTNF-a). Methods: Reverse transcription polymerase chain reaction (RT-PCR) was used to amplify the TNFR-1 gene cDNA. The cell line T-T was transfected with the TNFR-1 gene and TNFR-1 protein activity was confirmed by Western blot. MTT assay was used to explore the effect and mechanism of T-T cell line treated with rhTNF-a. Agarose gel electrophoresis and electron microscopy were used to study cellular changes. Results: The eukaryotic expression plasmid pcDNA3-TNFR1 and the T-T cell line were constructed successfully. Compared with the parental Tca-8113 and T-pc cells transfected with pcDNA3 vector, the T-T cell line showed more obvious cytotoxic effects when treated with rhTNF-a. Conclusion: The combination of TNFR-1 gene transfection and rhTNF-a could effectively induce T-T cell apoptosis, thus presenting a novel experimental basis for human tongue carcinoma gene therapy.



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