Estrogen plays a critical role in AAV2-mediated gene transfer in ovarian cancer.
OBJECTIVE
The aim of our study was to develop an effective gene delivery system for ovarian cancer gene therapy.


METHODS
The expression of heparin sulfate proteoglycan (HSPG) and integrins alpha(upsilon)beta(3) and alpha(upsilon)beta(5) were analyzed with flow cytometry on 2 human ovarian cancer cell lines (OVCAR-3 and SKOV-3ip). The gene transduction efficiencies were evaluated with recombinant adeno-associated viral vector (rAAV)2-green fluorescent protein or rAAV2-lactase Z followed by flow cytometry or cytohistochemistry staining. The effect of 17beta-estradiol on ovarian cancer cell proliferation, HSPG, the expressions of integrins alpha(upsilon)beta(3) and alpha(upsilon)beta(5), and adeno-associated viral vector (AAV)2-mediated gene transduction were determined.


RESULTS
In the present study, we found: (1) a variation in HSPG and the expressions of integrins alpha(upsilon)beta(3) and alpha(upsilon)beta(5) between OVCAR-3 and SKOV-3ip; (2) that 17beta-estradiol was shown to significantly stimulate cell proliferation and integrin beta(5) expression in certain ovarian cancer cell lines; and (3) integrintargeted A520/N584RGD-rAAV2, which has alternative interactivity with integrins and abrogates the binding capacity HSPG, showed much higher gene transduction efficiency in ovarian cancer cells than rAAV2 in the presence/absence of 17beta-estradiol. Moreover, this RGD-modified rAAV2 exerted more efficient transduction in ovarian cancer cells in response to 17beta-estradiol.


CONCLUSIONS
Our findings implied that A520/N584RGD-rAAV2 may offer great potential for ovarian cancer treatment in vivo.
Shi
Wen-fang
WF
Laboratory of Gene and Viral Therapy, Eastern Hepatobiliary Surgical Hospital, Second Military Medical University, Shanghai 200433, China. shiwenfang1@gmail.com
Bartlett
Jeffrey S
JS
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D016923:Cell Death / Q000502:physiology; D045744:Cell Line, Tumor / Q000187:drug effects; D000229:Dependovirus / Q000235:genetics / Q000378:metabolism; D004958:Estradiol / Q000378:metabolism*; D005260:Female; D015774:Ganciclovir / Q000494:pharmacology; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006493:Heparin / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000235:genetics / Q000378:metabolism; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000628:therapy; D011509:Proteoglycans / Q000378:metabolism; D019030:Receptors, Vitronectin / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic
D000998:Antiviral Agents; D039302:Integrin alphaVbeta3; D011509:Proteoglycans; D019030:Receptors, Vitronectin; C036090:heparin proteoglycan; C064660:integrin alphaVbeta5; D004958:Estradiol; D006493:Heparin; D015774:Ganciclovir