简介

The recombinant retrovirus vector pLXSN 16 E6/E7 containing the HPV-16 E6/E7 genes was used to transfect the ectotropic packaging cell line Psi-2.Virus produced by the Psi-2 cells was used to infect the amphotropic packaging cell line PA317.Virus produced by the PA317 cells was used to transduce primary PTCs.Although pLXSN 16 E6/E7 also confers resistance to neomycin, selection in G418 was not used to isolate transduced clones.The cell line appears to be derived from a single cell based on Southern and FISH analysis.The E6/E7 genes are present in the HK-2 genome as determined by PCR.The cells retain a phenotype indicative of well differentiated PTCs.They are positive for alkaline phosphatase, gamma glutamyltranspeptidase, leucine aminopeptidase, acid phosphatase, cytokeratin, alpha 3,beta 1 integrin, and fibronectin.The cells are negative for factor VIII related antigen, 6.19 antigen and CALLA endopeptidase.HK-2 cells retain functional characteristics of proximal tubular epithelium such as Na+ dependent / phlorizin sensitive sugar transport and adenylate cyclase responsiveness to parathyroid, but not to antidiuretic hormone.The cells are capable of gluconeogenesis as evidenced by their ability to make and store glycogen.HK-2 cells are anchorage dependent.The cells will not grow in methylcellulose, soft agar or suspension.HK-2 cells can reproduce experimental results obtained with freshly isolated PTCs.

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