The Eukaryotic Expression Of HHV-8 K13 And Its Effect On The Proliferation In Human Endothelial And Prostate Cancer Cells

Background and objective: So far, close associations were proved between cervix cancer and human papillomavirus or nasopharyngeal carcinoma and Epstein-Barr virus. Recently, several studies have focused on the relationship between some kind of virus infection and the development and progression of proatate cancer, in particular human herpesvirus 8 (HHV-8). HHV-8, called Kaposi's sarcoma-associated herpesvirus (KSHV), was first discovered in lesion tissue of acquired immunodeficiency syndrome-related Kaposi's sarcoma (AIDS-KS) in 1994. Viral FLICE inhibitory protein (v-FLIP) gene, named K13 gene, is one of latent infection gene in HHV-8. K13 gene plays different roles in different kinds of cells. We tried to clone K13 gene and observe its effects to the growth in human endothelial and prostate cancer cellsMethods: We constructed the recombinant plasmid PEF-K13-Puro, which was identified with enzyme digestion and nucleotide sequences analysis; the recombinant plasmid PEF-K13-Puro was transiently trancfected into EA.HY926 cells and PC-3 cells. The expression of K13 protein in both cells was analyzed by Western blot. The function of K13 protein in cell cycle and cell proliferation were measured by flow cytometry and MTT assay, respectively.Results: Nucleotide sequences analysis indicated that the cloned K13 sequence was 100% homology with K13 gene previously registered in GenBank. The specific band of K13 protein was detected at the expected place by Western blot. In addition, the results of the flow cytometry and MTT assay showed that the HHV-8 K13 protein could significantly inhibit the proliferation in human endothelial and prostate cancer cells.Conclusions: Finally, we successfully cloned HHV-8 K13 gene, whose protein might inhibit the growth of human endothelial and prostate cancer cells.