title: Influence of transfection with connexin 26 gene on malignant potential of human hepatoma cells.
abstract: We investigated the effect of transfection with connexin (Cx) 26 gene on the malignant potential of PLC/PRF/5 hepatoma cells, observing changes in their morphological features, alpha-fetoprotein (AFP) expression, cell proliferation and apoptosis in vitro, and their tumor growth in vivo. Fluorescence-activated cell sorting (FACS) analysis showed that 10.6% of PLC/PRF/5 hepatoma cells transfected with Cx26 cDNA expressed excessive Cx26, and the spread of lucifer yellow was wider in the colony of stable transfectants (PLC/Cx26) after its microinjection than in control. Nucleo-cytoplasmic (N/C) ratio was significantly lower in PLC/Cx26 (P < 0.0001). Cell proliferation assay showed significantly lower numbers in PLC/Cx26 on day 10 after seeding than in control (P = 0.0039), and AFP level /10(5) cells was significantly lower in medium of PLC/Cx26 (P = 0.0039). The number of proliferating cell nuclear antigen (PCNA)-positive cells was less in PLC/Cx26 in vitro than in control (P = 0.0039), and single-stranded DNA (ssDNA)-positive cells were more abundant in the colony of PLC/Cx26 (P = 0.029). Tumor volume in SCID mice was significantly smaller in the group of PLC/Cx26 than in the control (P < 0.01) throughout the observation period, and tumor weight of PLC/Cx26 was significantly lower (P = 0.0019) week 9 after inoculation. Transfection with Cx26 cDNA inhibited dedifferentiation, suppressed cell proliferation, and apoptosis was induced. Tumor growth of PLC/Cx26 was retarded. These findings suggest that transfection with Cx26 gene into human hepatoma cells reduces their malignant potential.
authors.affiliation: Third Department of Internal Medicine, Kyoto Prefectural University of Medicine, Kamigyo-ku, Kyoto 602-8566, Japan. amura@sun.kpu-m.ac.jp
authors.forename: Akira
authors.initials: A
authors.lastname: Muramatsu
authors.forename: Masaki
authors.initials: M
authors.lastname: Iwai
authors.forename: Teruhisa
authors.initials: T
authors.lastname: Morikawa
authors.forename: Saiyu
authors.initials: S
authors.lastname: Tanaka
authors.forename: Takahiro
authors.initials: T
authors.lastname: Mori
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Harada
authors.forename: Takeshi
authors.initials: T
authors.lastname: Okanoue
mesh_terms: D000818:Animals; D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D002455:Cell Division; D002467:Cell Nucleus / Q000378:metabolism; D002469:Cell Separation; D002478:Cells, Cultured; D000072259:Connexin 26; D017630:Connexins / Q000235:genetics*; D003593:Cytoplasm / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism; D005434:Flow Cytometry; D017629:Gap Junctions / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D016513:Mice, SCID; D009362:Neoplasm Metastasis; D010957:Plasmids / Q000378:metabolism; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured; D000509:alpha-Fetoproteins / Q000378:metabolism
chemical_list: D017630:Connexins; D018076:DNA, Complementary; D004277:DNA, Single-Stranded; C000607042:GJB2 protein, human; C000607043:Gjb2 protein, mouse; D018809:Proliferating Cell Nuclear Antigen; D000509:alpha-Fetoproteins; D000072259:Connexin 26