HepaCAM Inhibited Proliferation through a Wnt/β-catenin Pathway in Human Bladder Cancer Cell T24

Abstract: To investigate the effect of HepaCAM on proliferation and Wnt/β-catenin pathway in bladder cancer cells， T24 cells were done with blank treatment， Ad-GFP treatment and Ad-GFP-HepaCAM treatment， respectively. Then， CCK-8 assay was used to measure the effect of HepaCAM on the ability of cellular proliferation， the effects of HepaCAM on protein and mRNA expressions of β-catenin， c-Myc and cyclinD1 were evaluated by Western blot and qRT-PCR. In addition， T24 cells were treated with Wnt/β-catenin activator LiCl. MTT was used to detect cell growth， the phosphorylation level of GSK3β(try216) and the protein level of β-catenin， c-Myc and cyclinD1 were examined by Western blot， and the ability of cell colony was assessed by colony formation assay. Our research indicated that overexpression of HepaCAM could suppress cell growth and down-regulate the expression of β-catenin， c-Myc and cyclinD1 at mRNA and protein levels. Meanwhile， after T24 cells were treated with 5， 10， 20 μmol/L of LiCl for 2 h， cells proliferation was promoted and 10 μmol/L of LiCl could decrease the phosphorylation level of GSK3β， leading to activation of Wnt/β-catenin pathway. Moreover， combination treatment of 10 μmol/L of LiCl and Ad-GFP-HepaCAM could reverse the inhibition effect of HepaCAM on protein expression levels of β-catenin， c-Myc and cyclinD1 as well as cells growth. In conclusion， our study showed that expression of Hepa-CAM suppressed proliferation through a Wnt/β-catenin signaling pathway in bladder cancer cell T24.

CSTR: 32200.14.cjcb.2015.05.0012