1998 Fiscal Year Final Research Report Summary
Cloning of promoter of squamous cell carcinoma antigen and tissue secific gene therapy for cervical cancer
| Project/Area Number |
09671688
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | Ehime University |
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Principal Investigator |
HAMADA Katsuyuki Ehime University, School of Medicine, Obstetrics and Gynecology, Instructor, 医学部附属病院, 講師 (90172973)
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| Project Period (FY) |
1997 – 1998
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| Keywords | cervical cancer / squamous cell carcinoma / promoter / gene therapy / tissue specific / squamous cell carcinoma antigen 1 / squamous cell carcinoma antigen 2 / transcription regulation |
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| Research Abstract |
To select the suitable cells for promoter assay, expression of RNA arid protein of squamous cell carcinoma antigen (SCCA) 2 was studied in 40 human cell lines including cervical, skin and other squamous cell carcinoma cell lines, and endometrial, ovarian and other adenocarcinoma cell lines, and normal keratiocytes. ELIZA was used to detect SCCA protein expression in supernatant of each cell. This ELIZA did not differentiate SCCA1 and SCCA2 because of high homology of these 2 proteins. RT-PCR was used to detect SCCA2 RNA expression in each cell. Northern blot analysis was not used because of high homology (98%) of DNAs between SCCA1 and SCCA2. SCCA protein was detested in the supernatant of each squamous cell carcinoma cell line ; 18 ng/ml in SKG llla cells, 1 ng/ml in normal human keratinocyte, NHK, and <0.01 ng/ml in all adenocarcinoma cells. DNA-PCR analysis showed the clear bands of cDNAs of SCCA 1 and SCCA2 with high specificity from 1O^2 to 10^9 copies. RT-PCR analysis demonstrated the results which was well correlated with those of the ELIZA and adenocarcinoma cells did not show any bands. Transcription start site was 9bp upstream from the first codon, which was determined by SMART system. Deletion assay demonstrated the high promoter activity in the 250-500 bp upstream from the first codon. This promoter activity is the most in the SKG llla cells, 10% of that of SKGllla in the NHK, and absent in the SKOV3 cells. Promoter activity of SCCA2 is highly specific in the squamous cell carcinoma and promissing for the tissue specific gene therapy for squamous cell carcinoma.
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[Publications] Gotoh,A.,Kao,C.,Ko,S.C.,Hamada,K.,Roth,J.A.Chung,L.W.K.: "Gene therapy with p21WAF1/CIP1 adenovirus for human prostate cancers." Journal of Urology.158. 636-641 (1997)
Description
「研究成果報告書概要(和文)」より
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[Publications] Bouvet,M.,Fang,B,Ekmekcioglu,SJiL,Bucana,CD Hamada,K,Grimm,EA Roth,JA.: "Suppression of the immune response to an adenovirus vector and enhancement of intratumoral transgene expression by low-dose etoposide." Gene Therapy. 5. 189-195 (1998)
Description
「研究成果報告書概要(和文)」より
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[Publications] Kihana,T,Fujioka,T Hamada,K,Kito,K Takahashi,A,Tsukayama,C,Ito.M.: "Association of replication error positive phenotype with lymphocyte infiltration in endometrial cancers." Japanese Journal of Cancer Research. 89. 895-902 (1998)
Description
「研究成果報告書概要(和文)」より
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[Publications] Sarkar,A,Mitchell,MF,Hamada,K,Buchl,SJ Satterfield,WC Schapro,SJ Keeling,MF,Sastry,KJ.: "Evaluation of cellular immune response in rhesus monkeys subjected to adenovirus-mediated gene transfer into cervix." Cancer Gene Therapy.in print.
Description
「研究成果報告書概要(和文)」より
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[Publications] Gotoh, A., Kao, C., Ko, S.C., Hamada, K., Roth, J.A.Chung, L.W.K.: "Gene therapy with p21WAF1/CIP1 adenovirus for human prostate cancers." Journal of Urology. 158. 636-641 (1997)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Bouvet, M., Fang, B, Ekmekcioglu, SJiL, Bucana, CD Hamada, K, Grimm, EA Roth, JA.: "Suppression of the imOmune response to an adenovirus vector and enhancement of intratumoral transgene expression by low-dose etoposide." Gene Therapy. 5. 189-195 (1998)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Kihana, T, Fujioka, T Hamada, K, Kito, K, Takahashi, A, Tsukayama, C, Ito.M.: "Association of replication error positive phenotype with lymphocyte infiltration in endometrial cancers." Jpn.J.Cancer Res.89. 895-902 (1998)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Sarkar, A, Mitchell, MF, Hamada, K, Buchl, SJ Satterfield, WC Schapiro, SJ Keeling, ME,Sastry, KJ.: "Evaluation of cellular immune response in rhesus monkeys subjected to adenovirus-mediated gene transfer into cervix." Cancer Gene Ther.(in print).
Description
「研究成果報告書概要(欧文)」より
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