Masatoshi Tagawa

Publications (2)4.55 Total impact

• Article: Adenovirus vectors with chimeric type 5 and 35 fiber proteins exhibit enhanced transfection of human pancreatic cancer cells.

  Eiji Toyoda, Ryuichiro Doi, Kazuhiro Kami, Tomohiko Mori, Daisuke Ito, Masayuki Koizumi, Atsushi Kida, Kazuyuki Nagai, Tatsuo Ito, Toshihiko Masui, Michihiko Wada, Masatoshi Tagawa, Shinji Uemoto
  [show abstract] [hide abstract]
  ABSTRACT: Adenovirus (Ad) vectors are widely used for gene transfer. Efficient gene transfer into malignant cells is an important requirement for anticancer gene therapy, but transgene expression after transfer with adenoviral vectors varies among different cancer cell lines. Recently, Ad vectors containing chimeric type 5 and 35 fiber proteins have been developed. We evaluated the expression of coxsackie and adenovirus receptor (CAR), as well as integrins alphaV, beta3 and beta5, in seven human pancreatic cancer cell lines and assessed the relationship between expression of these molecules and Ad transfection efficiency. We compared the transfection efficiency of a conventional type 5 Ad vector (Ad5GFP) with that of an Ad vector containing chimeric type 5 and 35 fiber proteins (Ad5/35GFP), which expressed green fluorescent protein (GFP) driven by the cytomegalovirus promoter. There was strong CAR expression by AsPC-1, CFPAC-1 and PANC-1 cells, whereas the other cell lines showed weak expression. There was strong integrin beta3 expression by MIAPaCa-2, PANC-1 and Suit-2 cells, but expression by AsPC-1, BxPC-3, CFPAC-1 and HPAC cells was weak. Transfection efficiency of the vectors for human pancreatic cancer cell lines was not directly related to the CAR or integrin expression. However, transfection by Ad5/35GFP was significantly greater than by Ad5GFP at MOIs of 10 and 25 in all five human pancreatic cell lines. In conclusion, the Ad5/35GFP vector mediates more efficient gene transfer to human pancreatic cancer cells. These results may have implications for improving the efficiency of Ad-mediated gene transfer and developing adenoviral vectors.
  International Journal of Oncology 01/2009; 33(6):1141-7. · 2.40 Impact Factor
• Source

  Available from: PubMed Central

  Article: Midkine promoter-based conditionally replicative adenovirus therapy for midkine-expressing human pancreatic cancer.

  Eiji Toyoda, Ryuichiro Doi, Kazuhiro Kami, Tomohiko Mori, Daisuke Ito, Masayuki Koizumi, Atsushi Kida, Kazuyuki Nagai, Tatsuo Ito, Toshihiko Masui, Michihiko Wada, Masatoshi Tagawa, Shinji Uemoto
  [show abstract] [hide abstract]
  ABSTRACT: To develop a novel therapeutic strategy for human pancreatic cancer using a midkine promoter-based conditionally replicating adenovirus. 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. 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. 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.
  Journal of Experimental & Clinical Cancer Research 02/2008; 27:30. · 2.15 Impact Factor