title: Growth inhibition of human pancreatic cancer cells by human interferon-beta gene combined with gemcitabine.
abstract: We examined the anti-tumor effect of cationic multilamellar liposome containing human IFN-beta (huIFN-beta) gene against cultured human pancreatic cancer cells. We also evaluated the combined effect of huIFN-beta gene entrapped in liposomes and gemcitabine. Furthermore, we examined the anti-tumor mechanisms of the therapy, with emphasis on the Ras-related signal pathway. Three human pancreatic cancer cell lines (AsPc-1, MIAPaCa-2, and PANC-1) were used in this study. The growth inhibition together with the therapy were evaluated by WST-1 assay; the production of huIFN-beta protein was measured by ELISA; the cell cycle and apoptosis were analyzed using a FACScan flow cytometer; the protein levels of Son of sevenless (SOS-1) and Ras-GAP were measured by Western blotting; and the activation of Ras-GTP was evaluated by the immunoprecipitation method. As a result, we found that huIFN-beta gene entrapped in liposomes demonstrated a strong anti-tumor effect against human pancreatic cancer cells. The treatment that combined huIFN-beta gene entrapped in liposomes and gemcitabine was more effective than each treatment alone. Although gemcitabine remarkably reduced the level of SOS-1, the above combined therapy reduced the level of SOS-1 even more significantly. Both huIFN-beta gene entrapped in liposomes and the com-bination of huIFN-beta gene entrapped in liposomes and gemcitabine increased the level of Ras-GAP, and decreased the activity of Ras-GTP. These results suggest that this combination therapy can induce strong anti-tumor activity against human pancreatic cancer cells through the regulation of the Ras-related signal pathway.
authors.affiliation: Second Department of Surgery, Toyama Medical and Pharmaceutical University, Nagoya, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Endou
authors.forename: Masaaki
authors.initials: M
authors.lastname: Mizuno
authors.forename: Takuya
authors.initials: T
authors.lastname: Nagata
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Tsukada
authors.forename: Norimoto
authors.initials: N
authors.lastname: Nakahara
authors.forename: Takaya
authors.initials: T
authors.lastname: Tsuno
authors.forename: Hirokatsu
authors.initials: H
authors.lastname: Osawa
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Kuno
authors.forename: Mitsugu
authors.initials: M
authors.lastname: Fujita
authors.forename: Manabu
authors.initials: M
authors.lastname: Hatano
authors.forename: Jun
authors.initials: J
authors.lastname: Yoshida
mesh_terms: D017304:Annexin A5 / Q000494:pharmacology; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D017209:Apoptosis; D015153:Blotting, Western; D002412:Cations; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002469:Cell Separation; D003131:Combined Modality Therapy; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D047468:Immunoprecipitation; D016899:Interferon-beta / Q000235:genetics* / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000473:pathology* / Q000628:therapy*; D010957:Plasmids / Q000378:metabolism; D020837:SOS1 Protein / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D020703:ras GTPase-Activating Proteins / Q000378:metabolism; D000093542:Gemcitabine
chemical_list: D017304:Annexin A5; D000964:Antimetabolites, Antineoplastic; D002412:Cations; D017077:Culture Media, Conditioned; D008081:Liposomes; D020837:SOS1 Protein; D020703:ras GTPase-Activating Proteins; D003841:Deoxycytidine; D016899:Interferon-beta; D000093542:Gemcitabine