Short hairpin RNA targeting beta-catenin suppresses cell proliferation and induces apoptosis in human gastric carcinoma cells.
OBJECTIVE
Aberrant activation of Wnt/beta-catenin signaling is involved in various cancers, including human gastric cancer. Here we investigate the role of Wnt/beta-catenin signaling in regulating gastric cancer cell apoptosis.


METHODS
Expression of beta-catenin was investigated after transfection with beta-catenin short hairpin RNA (shRNA) in gastric cancer cells by Western blotting and immunofluorescence analysis. beta-catenin/T-cell factor transcriptional activity was also investigated by using a luciferase reporter assay. Next, the effects of beta-catenin shRNA on cell proliferation and apoptosis were evaluated by the 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide assay and flow cytometric analysis. To investigate the precise mechanism of these effects, a comprehensive analysis was performed using a cDNA microarray.


RESULTS
shRNA targeting beta-catenin resulted in a significant decrease in beta-catenin expression, and its nuclear localization and cell proliferation. Meanwhile, increased cell apoptosis was confirmed. The comprehensive analysis showed that shRNA targeting beta-catenin upregulated 26 apoptosis-related genes (including PERP, TRAF3, PDCD2, TNFRSF25, AKT2 and YWHAZ) and downregulated 48 apoptosis-related genes (including MALT1, IRAK1, TNFAIP3, PPP1R13L, TRIP and YWHAB) in gastric cancer cells. Pathway analysis suggested that the nuclear factor-kappaB pathway was involved in beta-catenin knockdown-induced apoptosis.


CONCLUSIONS
Attenuation of beta-catenin by shRNA resulted in suppressed cell proliferation and apparent apoptosis, suggesting that beta-catenin may be a target for therapy of gastric cancer.
Jiang
Hong
H
Department of Gastrointestinal Surgery, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, PR China.
Xia
Jianguo
J
Kang
Jian
J
Ding
Yongbin
Y
Wu
Wenxi
W
D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D014408:Biomarkers, Tumor / Q000235:genetics; D015153:Blotting, Western; D002277:Carcinoma / Q000235:genetics / Q000378:metabolism* / Q000628:therapy; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015536:Down-Regulation; D005455:Fluorescent Antibody Technique; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D009154:Mutation; D016328:NF-kappa B / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D034741:RNA, Small Interfering / Q000378:metabolism* / Q000627:therapeutic use; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013274:Stomach Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000628:therapy; D014178:Translocation, Genetic; D015854:Up-Regulation; D051176:beta Catenin / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*
D014408:Biomarkers, Tumor; C495270:CTNNB1 protein, human; D016328:NF-kappa B; D034741:RNA, Small Interfering; D051176:beta Catenin