Midkine promoter-based conditionally replicative adenovirus therapy for midkine-expressing human pancreatic cancer
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- 作者：Eiji Toyoda (1)
Ryuichiro Doi (1)
Kazuhiro Kami (1)
Tomohiko Mori (1)
Daisuke Ito (1)
Masayuki Koizumi (1)
Atsushi Kida (1)
Kazuyuki Nagai (1)
Tatsuo Ito (1)
Toshihiko Masui (1)
Michihiko Wada (1)
Masatoshi Tagawa (2)
Shinji Uemoto (1) - 刊名：Journal of Experimental & Clinical Cancer Research
- 出版年：2008
- 出版时间：December 2008
- 年：2008
- 卷：27
- 期：1
- 全文大小：2174KB
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Ryuichiro Doi (1)
Kazuhiro Kami (1)
Tomohiko Mori (1)
Daisuke Ito (1)
Masayuki Koizumi (1)
Atsushi Kida (1)
Kazuyuki Nagai (1)
Tatsuo Ito (1)
Toshihiko Masui (1)
Michihiko Wada (1)
Masatoshi Tagawa (2)
Shinji Uemoto (1)
1. Department of Hepato-Biliary-Pancreatic Surgery and Transplantation, Kyoto University, Japan
2. Division of Pathology, Chiba Cancer Center Research Institute, Chiba, Japan - ISSN：1756-9966
文摘
Background To develop a novel therapeutic strategy for human pancreatic cancer using a midkine promoter-based conditionally replicating adenovirus. Methods We examined midkine mRNA expression and midkine protein expression by seven human pancreatic cancer cell lines (AsPC-1, BxPC-3, CFPAC-1, HPAC, MIAPaCa-2, PANC-1, and Suit-2), as well as by non-cancerous pancreatic tissue and pancreatic cancers. Midkine promoter activity was measured in cancer cell lines by the dual luciferase reporter assay. Adenoviral transduction efficiency was assessed by fluorescent staining of cancer cell lines using adenovirus type 5 containing the green fluorescent protein gene (Ad5GFP). Replication of adenovirus type 5 containing the 0.6 kb midkne promoter (Ad5MK) was assessed by the detection of E1 protein in cancer cell lines. The cytotoxicity of Ad5MK for cancer cells was evaluated from the extent of growth inhibition after viral infection. Infection and replication were also assessed in nude mice with subcutaneous Suit-2 tumors by intratumoral injection of Ad5MK, Ad5GFP, or vehicle. E1a mRNA expression in the treated tumors and expression of the replication-specific adenoviral hexon protein were evaluated. Finally, the anti-tumor activity of Ad5MK against intraperitoneal xenografts of Suit-2 pancreatic cancer cells was examined after intraperitoneal injection of the virus. Results Both midkine mRNA expression and midkine protein expression were strong in AsPC-1 and CFPAC-1 cell liens, moderate in BxPC-3, HPAC, and Suit-2 cell lines, and weak in PANC-1 and MIAPaCa-2 cell lines. Expression of midkine mRNA was significantly stronger in pancreatic cancers than in non-cancerous pancreatic tissues. The relative luciferase activity mediated by the 0.6 kb midkne fragment in AsPC-1, PANC-1, and Suit-2 cell lines was approximately 6 to 20 times greater than that in midkne-negative MIAPaCa-2 cell lines. Pancreatic cancer cell lines exhibited a heterogeneous adenoviral transduction profile. E1A expression was higher in cell lines with strong midkine expression than in cell lines with weak midkine expression. Ad5MK showed much greater cytotoxicity for midkine-expressing Suit-2 and PANC-1 cell lines than for midkine-negative MIAPaCa-2 cell lines. In the Suit-2 subcutaneous xenograft model, expression of E1A was detected in Ad5MK-treated tumors, but not in untreated and Ad5GFP-treated tumors. In the Suit-2 intraperitoneal xenograft model, the Ad5MK group survived for significantly longer than the Ad5GFP, PBS, and untreated groups. Conclusion Ad5MK has an anti-tumor effect against human pancreatic cancer cell lines that express midkine mRNA. Midkine promoter-based conditionally replicative adenovirus might be a promising new gene therapy for pancreatic cancer.