title: Construction of recombinant human IFNα-2b BCG and its antitumor effects on bladder cancer cells in vitro.
abstract: We constructed recombinant Bacille Calmette-Guérin (rBCG) that secreted human interferon alpha 2b (hIFNα-2b), and investigated its antitumor effects on bladder cancer cells in vitro. The recombinant plasmid phIFN-α-2b was constructed using pMAO-4 and transformed into BCG. The supernatant was collected at various times and IFN-γ, interleukin (IL)-12, and tumor necrosis factor (TNF)-α were detected using an enzyme-linked immunosorbent assay. EJ cells were cultivated for 24, 48, and 72 h, together with rBCG, wild-type BCG (wBCG), or wBCG+IFN-α-2b. rBCG capable of secreting cytokine IFNα-2b was constructed. On the 4th day of culture, the IFNα-2b secreted by rBCG reached a maximum. wBCG and rBCG showed no significant difference on cell growth rate over 7 days of incubation in 7H9 medium. wBCG and rBCG were both positive for acid-fast staining, and showed mycobacterial characteristics of intercellular connection in clusters with no clear abnormalities. Higher levels of IFN-γ, TNF-α, and IL-12 were induced by rBCG compared with wBCG or MAO4-rBCG (P < 0.05). rBCG may induce lymphocyte proliferation; the proliferation ratio was higher than those induced by wBCG and wBCG+IFN. rBCG had direct anti-proliferative effects on EJ cells. An MTT assay showed that rBCG inhibited the proliferation of bladder cancer cells and had more activity compared with wBCG (P < 0.05). The highest anti-tumor activity of lymphocytes was stimulated by rBCG (20.31-51.22%). rBCG-IFNα-2b induces enhanced cytotoxicity against bladder cancer cells in vitro and may be used as an alternative to BCG for bladder cancer patients.
authors.affiliation: Department of Urology, The Second Hospital of Tianjin Medical University, Tianjin Institute of Urology, Tianjin, China.
authors.forename: E
authors.initials: E
authors.lastname: Sun
authors.affiliation: Department of Urology, The Second Hospital of Tianjin Medical University, Tianjin Institute of Urology, Tianjin, China.
authors.forename: X
authors.initials: X
authors.lastname: Nian
authors.affiliation: Department of Urology, The Second Hospital of Tianjin Medical University, Tianjin Institute of Urology, Tianjin, China.
authors.forename: C
authors.initials: C
authors.lastname: Liu
authors.affiliation: Tianjin Central Hospital of Gynecology Obstetrics, Tianjin, China.
authors.forename: X
authors.initials: X
authors.lastname: Fan
authors.affiliation: Department of Urology, The Second Hospital of Tianjin Medical University, Tianjin Institute of Urology, Tianjin, China hanruifa2013@sina.com.
authors.forename: R
authors.initials: R
authors.lastname: Han
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis; D002295:Carcinoma, Transitional Cell / Q000276:immunology / Q000628:therapy*; D045744:Cell Line, Tumor / Q000187:drug effects; D049109:Cell Proliferation; D002470:Cell Survival / Q000187:drug effects; D016207:Cytokines / Q000378:metabolism; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D007155:Immunologic Factors / Q000494:pharmacology; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000494:pharmacology*; D009163:Mycobacterium bovis / Q000276:immunology* / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D018417:Th1 Cells / Q000378:metabolism / Q000502:physiology; D001749:Urinary Bladder Neoplasms / Q000276:immunology / Q000628:therapy*
chemical_list: D000970:Antineoplastic Agents; D016207:Cytokines; D007155:Immunologic Factors; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins