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[Establishment of human immortalized oral epithelial cell line HIO615 induced by HPV16 E6 and E7]
Zhiyuan Zhang1, Patima Sdek, Jun Cao
1Department of Oral and Maxillofacial Surgery, School of Stomatology, Shanghai Second Medical University, Shanghai 200011, China.
Objective:
To establish an immortalized oral epithelial cell line.
Methods:
Normal human oral epithelial cells were transfected with HPV16E6/E7 open reading frames using recombinant retroviral system pLXSN. Expression of HPV16E6 and E7 protein were tested by Western blot in three kinds of cells. To define cellular biological characterization of HPV16E6/E7 transfected cells, a series analysis were performed, including protraction of growth curve, HE staining, immunocytochemical staining and scanning electron microscope observation. The tumorigenicity was assessed by colony formation and transplanting the cells into nude mice.
Results:
Human oral epithelial cells transfected with HPVE6/E7 has been in culture for over 18 months. The cell line was named HIO615. Western blot analysis showed HIO615 expressed HPV16 E6 and E7 protein. HIOC were positive for cytokeratin, tonofibril and desmosome as observed by scanning electron microscope. The number of large colonies of dense multilayer cells was low (0.77%). No tumor developed in nude mice injected subcutaneously with HIOEC.
Conclusion:
A human immortalized oral epithelial cell line induced by HPV16E6 and E7 has been successfully established.