Tumor-specific Gene Therapy for Renal Cell Carcinoma Using MN/CA9-directed Replication-competent Adenovirus.
- Author:
Se Joong KIM
1
;
Miwon AHN
;
Ho Yeong LIM
;
Cheol Hyun CHUNG
;
Thomas A GARDNER
;
Chinghai KAO
;
Sang Jin LEE
;
Min Kyu CHOI
;
Young Soo KIM
Author Information
1. Department of Urology, Ajou University School of Medicine, Suwon, Korea.
- Publication Type:In Vitro ; Original Article
- Keywords:
Renal cell carcinoma;
Gene therapy;
Adenovirus;
Virus replication
- MeSH:
Adenoviridae*;
Blotting, Western;
Carcinoma, Renal Cell*;
Cell Line;
DNA, Complementary;
Genetic Therapy*;
Glycoproteins;
Humans;
Kidney;
Prognosis;
RNA, Messenger;
Virus Replication
- From:Korean Journal of Urology
2004;45(5):456-462
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: A new therapeutic approach is needed in patients with metastatic renal cell carcinoma (RCC) because of a dismal prognosis. MN/CA9 is a transmembrane glycoprotein that was first identified in the human cervical carcinoma cell line, HeLa. Since MN/CA9 protein is highly expressed in RCC tissues, but not in normal kidney, we constructed a tumor-specific replication-competent adenoviral vector utilizing MN/CA9 promoter (Ad-MN/CA9-E1a) and demonstrated its selective cytotoxicity toward MN/CA9-expressing RCC cells in vitro. MATERIALS AND METHODS: MN/CA9-positive (HeLa, SK-RC-52) and MN/ CA9-negative (SK-RC-29) cells were used. RT-PCR assay for MN/CA9 mRNA was performed in each cells. Ad5 E1a protein production in each cells after infection with Ad-MN/CA9-E1a was determined by western blot analysis. In vitro cytotoxicity assay was performed for assessing the selective cytotoxicity of Ad-MN/CA9-E1a to MN/CA9-expressing cells. RESULTS: RT-PCR assay showed that a distinct 255-bp fragment corresponding to the sequence within MN/CA9 cDNA was detected in HeLa and SK-RC-52 cells, but SK-RC-29 cells did not have MN/CA9 transcripts. Western blot analysis demonstrated that HeLa and SK-RC-52 cells showed much stronger Ad5 E1a protein expressions compared with SK-RC-29. In vitro cytotoxicity assay revealed that the growth of MN/CA9-positive cells was significantly inhibited with 0.1-1MOI of Ad-MN/CA9-E1a, but the growth of MN/CA9-negative cells (SK-RC-29) could only be inhibited by as many as 100MOI. CONCLUSIONS: These results suggest that a novel replication-competent adenoviral vector mediated by MN/CA9 promoter, Ad-MN/CA9-E1a, can selectively replicate in MN/CA9-expressing cancer cells with cytotoxic effects and may be utilized for the treatment of RCC.
