[Immortalization of human embryonic cervical epithelial cells induced by E6, E7 genes of human papillomavirus 16].
OBJECTIVE
To establish an immortalized cell line derived from the embryonic cervical epithelium by infection with the recombinant adeno-associated virus (rAAV) containing human papillomavirus (HPV)16 E6, E7, and to study the biological features of cervical cancer cell line.


METHODS
Human embryonic cervical tissues were cultured in keratinocyte free serum (K-FS) medium and infected with rAAV containing HPV16 E6, E7. Morphological features and growth rate were examined by light, electronic and fluorescence microscopies. The fragments of E6, E7 were detected by polymerase chain reaction (PCR) and laser confocal microscopy. The biological characteristics of human cervical epithelium were observed by soft agar culture, scid mice inoculation and chromosome analysis. Cell proliferative dynamics was plotted by flow cytometry.


RESULTS
After a long-term culture, the phenotype kept the characteristics of primary epithelial cells. They showed monolayer, anchorage-dependent and attachment-inhibited growth without forming colonies in soft agar culture. They were non-oncogenic when inoculated into scid mice. The tonofilament expression in the cervical cancer cells was inspected by electronic microscopy, demonstrating that the cells were squamous epithelium in origin. The cell line contained HPV16 E6, E7 genes by PCR and laser confocal detection. Chromosome analysis disclosed that the karyotype was diploid or polyploid. The 11th chromosome was assumed to be the integration site by rAAV containing HPV16 E6, E7.


CONCLUSIONS
Establishment of the immortalized cervical epithelial cell line by infection with rAAV containing HPV16 E6, E7, supports that HPV16 E6, E7 may be the primary etiology of cervical cancer. It will facilitate further research on the etiology and pathogenesis of cervical cancer.
Cao
Ze-Yi
ZY
Department of Obstetrics and Gynecology, First Hospital, Beijing University, Beijing 100034, China.
Zhao
Jian
J
Liao
Qin-Ping
QP
Yang
Yi-Shu
YS
Zhou
Ling
L
Zeng
Yi
Y
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D002584:Cervix Uteri / Q000166:cytology* / Q000821:virology*; D004847:Epithelial Cells / Q000166:cytology* / Q000821:virology; D005260:Female; D005333:Fetus; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000235:genetics; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins / Q000096:biosynthesis* / Q000235:genetics
C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16