Selective activation of cervical microvascular endothelial cells by human papillomavirus 16-e7 oncoprotein.
BACKGROUND
Human papillomavirus type 16 (HPV16) is strongly implicated in the etiology of cervical cancer, with the expression of HPV16-encoded E7 oncoprotein in infected epithelial cells contributing to their malignant transformation. Although nuclear E7 interacts with several nuclear targets, we have previously shown that extracellular E7 can cause suppression of immune cell function. Moreover, cervical microvascular endothelial (CrMVEn) cells treated with E7 increase their expression of adhesion molecules. High levels of some cytokines in serum and in cervicovaginal secretions are associated with the progression of cervical cancer. In this study, we investigated the effects of extracellular E7 on cytokine production and on cytoskeleton structure of CrMVEn cells and vascular endothelial cells from different organs.


METHODS
Immunocytochemical staining and flow cytometry techniques were used to detect E7 in endothelial cells incubated with purified E7 protein. Laser scanning confocal microscopy was used to study the E7-induced modification of the endothelial cytoskeleton. An enzyme-linked immunosorbent assay was performed to measure the production of two cytokines, interleukin 6 (IL-6) and interleukin 8 (IL-8), by E7-treated endothelial cells. All statistical tests were two-sided.


RESULTS
Extracellular E7 was taken up by CrMVEn cells and localized to the cytoplasm. CrMVEn cells showed a statistically significant (P<.02) increase in the production of IL-6 and IL-8 after treatment with E7 compared with the controls. CrMVEn cells also produced higher levels of these cytokines than did the other endothelial cells (P<.01). E7 also induced marked alterations in the endothelial cytoskeleton of CrMVEn cells as a result of actin fiber polymerization.


CONCLUSIONS
These findings suggest a novel mechanism by which E7, as an extracellular factor, can play a role in the progression and dissemination of cervical cancer via its selective effects on endothelial cells.
D'Anna
R
R
Laboratoire de Physiologie Cellulaire, Université Pierre et Marie Curie, Paris, France.
Le Buanec
H
H
Alessandri
G
G
Caruso
A
A
Burny
A
A
Gallo
R
R
Zagury
J F
JF
Zagury
D
D
D'Alessio
P
P
D002448:Cell Adhesion; D002478:Cells, Cultured; D002584:Cervix Uteri / Q000098:blood supply* / Q000378:metabolism*; D003513:Cycloheximide / Q000494:pharmacology; D016207:Cytokines / Q000097:blood; D003593:Cytoplasm / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D015850:Interleukin-6 / Q000096:biosynthesis; D016209:Interleukin-8 / Q000096:biosynthesis; D008833:Microcirculation / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis*; D050725:Papillomavirus E7 Proteins; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D013997:Time Factors; D014471:Umbilical Veins / Q000166:cytology
D016207:Cytokines; D015850:Interleukin-6; D016209:Interleukin-8; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D011500:Protein Synthesis Inhibitors; D011994:Recombinant Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; D003513:Cycloheximide