BMC cancer · 2011
Loss of runt-related transcription factor 3 expression leads hepatocellular carcinoma cells to escape apoptosis.
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Nakanishi Yutaka, Shiraha Hidenori, Nishina Shin-ichi, Tanaka Shigetomi, Matsubara Minoru, Horiguchi Shigeru, Iwamuro Masaya, Takaoka Nobuyuki, Uemura Masayuki, Kuwaki Kenji, Hagihara Hiroaki, Toshimori Junichi, Ohnishi Hideki, Takaki Akinobu, Nakamura Shinichiro, Kobayashi Yoshiyuki, Nouso Kazuhiro, Yagi Takahito, Yamamoto Kazuhide
Аннотация
BACKGROUND: Runt-related transcription factor 3 (RUNX3) is known as a tumor suppressor gene for gastric cancer and other cancers, this gene may be involved in the development of hepatocellular carcinoma (HCC).
METHODS: RUNX3 expression was analyzed by immunoblot and immunohistochemistry in HCC cells and tissues, respectively. Hep3B cells, lacking endogenous RUNX3, were introduced with RUNX3 constructs. Cell proliferation was measured using the MTT assay and apoptosis was evaluated using DAPI staining. Apoptosis signaling was assessed by immunoblot analysis.
RESULTS: RUNX3 protein expression was frequently inactivated in the HCC cell lines (91%) and tissues (90%). RUNX3 expression inhibited 90±8% of cell growth at 72 h in serum starved Hep3B cells. Forty-eight hour serum starvation-induced apoptosis and the percentage of apoptotic cells reached 31±4% and 4±1% in RUNX3-expressing Hep3B and control cells, respectively. Apoptotic activity was increased by Bim expression and caspase-3 and caspase-9 activation.
CONCLUSION: RUNX3 expression enhanced serum starvation-induced apoptosis in HCC cell lines. RUNX3 is deleted or weakly expressed in HCC, which leads to tumorigenesis by escaping apoptosis.
METHODS: RUNX3 expression was analyzed by immunoblot and immunohistochemistry in HCC cells and tissues, respectively. Hep3B cells, lacking endogenous RUNX3, were introduced with RUNX3 constructs. Cell proliferation was measured using the MTT assay and apoptosis was evaluated using DAPI staining. Apoptosis signaling was assessed by immunoblot analysis.
RESULTS: RUNX3 protein expression was frequently inactivated in the HCC cell lines (91%) and tissues (90%). RUNX3 expression inhibited 90±8% of cell growth at 72 h in serum starved Hep3B cells. Forty-eight hour serum starvation-induced apoptosis and the percentage of apoptotic cells reached 31±4% and 4±1% in RUNX3-expressing Hep3B and control cells, respectively. Apoptotic activity was increased by Bim expression and caspase-3 and caspase-9 activation.
CONCLUSION: RUNX3 expression enhanced serum starvation-induced apoptosis in HCC cell lines. RUNX3 is deleted or weakly expressed in HCC, which leads to tumorigenesis by escaping apoptosis.
Предметные рубрики MeSH
AdolescentAdultAgedApoptosisApoptosis Regulatory ProteinsBcl-2-Like Protein 11Carcinoma, HepatocellularCaspasesCell Line, TumorCell ProliferationCell SurvivalCore Binding Factor Alpha 3 SubunitCulture Media, Serum-FreeGene Expression Regulation, NeoplasticHep G2 CellsHumansImmunoblottingImmunohistochemistryLiver NeoplasmsMembrane ProteinsMiddle AgedMutationProto-Oncogene ProteinsRNA InterferenceReverse Transcriptase Polymerase Chain ReactionYoung Adult