PMID: 20646651Jul 22, 2010Paper

Effect of siRNA-mediated beta-catenin gene on Wnt signal pathway in lung adenocarcinoma A549 cell

Zhonghua yi xue za zhi
Ying TengJian Wang

Abstract

To study the effects of siRNA-mediated beta-catenin down-regulation on Wnt/beta-catenin signal transduction pathway in lung adenocarcinoma A549 cells. A549 cell was divided into three groups: PGCsil-CTNNB1-siRNA group (transfection with beta-catenin interference plasmids), negative control group (transfection with negative control plasmids) and non-transfection group (blank control). For each group, real-time PCR was employed to detect the expression of beta-catenin and cyclin D1 (a target gene of Wnt/beta-catenin pathway). A MTT cell proliferation assay was performed to study the proliferating capacity. Flow cytometry was used for cell cycle analysis. Clone formation and Millicell chamber experiment were performed to evaluate the clone formation and migration capacities of cells. The expression of beta-catenin and cyclin D1 in PGCsil-CTNNB1-siRNA cell decreased at the mRNA level. It was lower than negative control (0.002+/-0.001 vs 0.023+/-0.002, P<0.01; 0.005+/-0.002 vs 0.040+/-0.020, P<0.05) and blank control groups (beta-catenin mRNA: 0.021+/-0.003, P<0.01; cyclin D1 mRNA: 0.042+/-0.004, P<0.05). Also the proliferating capacity at Days 5-7 was inhibited in comparison with negative control and blank control groups (P<0.05, P...Continue Reading

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