Evaluation of tissue-specific promoters in carcinomas of the cervix uteri.
BACKGROUND
Gene therapy is a novel approach for treatment of patients with advanced, recurrent, or metastatic cervical cancer. One effective way to direct transgene expression to specific tissues or tumors is the use of tissue-specific-promoters (TSPs). In the context of adenovirus (Ad)-mediated cancer gene therapy it is rational to choose a TSP which is highly expressed in the tumor but has potentially low activity in non-tumor cells, especially the liver. In this study, we have investigated several promoters which fulfill these criteria. Candidate cervical cancer specific TSPs include promoters of the genes for secretory leukoprotease inhibitor (SLPI), cyclooxygenase-2 (COX-2), Midkine (MK), vascular endothelial growth factor receptor type 1 (flt-1), vascular endothelial growth factor (VEGF), Survivin and the receptor for chemokine SDS-1 (CXCR4).


METHODS
To evaluate the specific gene expression of the different promoters in the context of cervical cancer, we constructed a panel of E1-deleted Ads that express luciferase under the control of the promoters of interest. We investigated various established cervical cancer cell lines, as well as purified primary cancer cells and normal control cells from the cervix uteri.


RESULTS
In all cell lines tested, promoters for MK, VEGF and CXCR4 showed the highest activity. Both MK and VEGF promoters also resulted in a high activity in primary cervical cancer cells. Interestingly, gene expression profiles correlate with luciferase activity in both cell lines and primary cancer samples.


CONCLUSIONS
Our study demonstrates that the promoters for MK and VEGF are active in cervical cancer. We believe that both promoters can be successfully employed as TSPs for gene therapy targeted to cervical cancer.
Rein
Daniel T
DT
Division of Human Gene Therapy, Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294-2172, USA.
Breidenbach
Martina
M
Nettelbeck
Dirk M
DM
Kawakami
Yosuke
Y
Siegal
Gene P
GP
Huh
Warner K
WK
Wang
Minghui
M
Hemminki
Akseli
A
Bauerschmitz
Gerd J
GJ
Yamamoto
Masato
M
Adachi
Yasuo
Y
Takayama
Koichi
K
Dall
Peter
P
Curiel
David T
DT
D000256:Adenoviridae / Q000235:genetics; D045744:Cell Line, Tumor; D051546:Cyclooxygenase 2 / Q000235:genetics; D016207:Cytokines / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000235:genetics; D005260:Female; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000235:genetics; D000078792:Midkine; D018995:Myosin Heavy Chains; D009363:Neoplasm Proteins / Q000235:genetics; D024501:Nonmuscle Myosin Type IIB; D011401:Promoter Regions, Genetic; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins / Q000235:genetics; D019718:Receptors, CXCR4 / Q000235:genetics; D053481:Secretory Leukocyte Peptidase Inhibitor; D000077022:Survivin; D002583:Uterine Cervical Neoplasms / Q000235:genetics*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics
C107489:BIRC5 protein, human; D016207:Cytokines; D016326:Extracellular Matrix Proteins; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D019718:Receptors, CXCR4; C506814:SLPI protein, human; D053481:Secretory Leukocyte Peptidase Inhibitor; C506815:Slpi protein, mouse; D000077022:Survivin; D042461:Vascular Endothelial Growth Factor A; D000078792:Midkine; D051546:Cyclooxygenase 2; D024501:Nonmuscle Myosin Type IIB; C103815:nonmuscle myosin type IIB heavy chain; D018995:Myosin Heavy Chains