Clone Of Human Xeroderma Pigmentosum Group D And Analysis Of Its Function In Melanoma Cells

BackgroundXeroderma pigmentosum group D (XPD) gene is the second subunit of basictranscript factor TFII H; it plays an important role in transcription and nucleotideexcision repair. Studies suggest that some genetic variants of the XPD gene may beassociated with risk of cutaneous melanoma.ObjectiveTo clone the cDNA sequences of human xeroderma pigmentosum group D andinvestigate the biological activity of the obtained XPD in melanoma cells.MethodsUsing the total RNA extracted from HeLa cells, we cloned the human full-lengthXPD by RT-PCR and inserted it into the pEGFP-N1plasmid vector. Then therecombinant plasmid pEGFP-N1/XPD was transfected into the human malignantmelanoma A375cells by the lipofectamine2000TM. The expression levels of XPD weredetected by Western blotting. The cell growth was examined by MTT..ResultsThe full-length cDNA sequences of XPD were obtained. The obtained cDNA was1234bp in size, which was consistent with the prediction. Green fluorescences wasobserved in the melanoma cells transfected with the recombinant plasmid，and theproliferation of the transfected cells was inhibited.ConclusionThe study provides evidence for the research of biological influence of XPD onmelanoma cells. The delivery of XPD may be a new useful strategy for treatment ofmelanoma or other tumor cells.