Journal Article

Forced expression of antisense 14-3-3β RNA suppresses tumor cell growth <i>in vitro</i> and <i>in vivo</i>

Akinori Sugiyama, Yohei Miyagi, Yuko Komiya, Nobuya Kurabe, Chifumi Kitanaka, Naoko Kato, Yoji Nagashima, Yoshiyuki Kuchino and Fumio Tashiro

in Carcinogenesis

Volume 24, issue 9, pages 1549-1559
Published in print September 2003 | ISSN: 0143-3334
Published online September 2003 | e-ISSN: 1460-2180 | DOI: https://dx.doi.org/10.1093/carcin/bgg113
Forced expression of antisense 14-3-3β RNA suppresses tumor cell growth in vitro and in vivo

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The 14-3-3 family proteins are key regulators of various signal transduction pathways including malignant transformation. Previously, we found that the expression of the 14-3-3β gene is deregulated as well as c-myc gene in aflatoxin B1 (AFB1)-induced rat hepatoma K1 and K2 cells. To elucidate the implication of 14-3-3β in tumor cell growth, in this paper we analyzed the effect of forced expression of antisense 14-3-3β RNA on the growth and tumorigenicity of K2 cells. K2 cells transfected with antisense 14-3-3β cDNA expression vector diminished their growth ability in monolayer culture and in semi-solid medium. Expression level of vascular endothelial growth factor mRNA was also reduced in these transfectants. Tumors that formed by the transfectants in nude mice were much smaller and histologically more benign tumors, because of their decreased level of mitosis compared with those of the parental cells. Frequency of apoptosis detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay was increased in the transfectant-derived tumors accompanying the inhibition of angiogenesis. In addition, over-expression of 14-3-3β mRNA was observed in various murine tumor cell lines. These results suggest that 14-3-3β gene plays a pivotal role in abnormal growth of tumor cells in vitro and in vivo.

Keywords: AFB1, aflatoxin B1; DMEM, Dulbecco's modified Eagle's medium; FCS, fetal calf serum; IGF-II, insulin-like-growth factor-II; INT, 1-p-iodophenyl-p-nitrophenyl-5-phenyltetrazolium chloride; ODNs, oligode­oxynucleotides; TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling; VEGF, vascular endothelial growth factor; vWF, von Willebrand factor

Journal Article.  7616 words.  Illustrated.

Subjects: Clinical Cytogenetics and Molecular Genetics

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