HepaCAM通过Wnt/β-catenin信号通路抑制膀胱癌细胞T24的增殖  

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

作　　者：万雪莲[1] 马菲菲[1] 马红莹 吕长坤[1] 

机构地区：[1]商丘医学高等专科学校医学检验技术教研室,商丘476100

出　　处：《中国细胞生物学学报》2015年第5期683-688,共6页Chinese Journal of Cell Biology

摘　　要：该研究探讨了肝细胞黏附分子(hepatocyte cell adhesion molecule,Hepa CAM)对膀胱癌细胞T24增殖的影响及其对Wnt/β-catenin信号通路的调控作用。T24细胞做空白处理、空载腺病毒(Ad-GFP)处理和Hepa CAM过表达腺病毒(Ad-GFP-Hepa CAM)处理,CCK-8法检测Hepa CAM对细胞增殖的影响,q RT-PCR和Western blot法检测Hepa CAM对β-catenin、c-Myc和cyclin D1的m RNA和蛋白表达水平的影响。采用Wnt/β-catenin信号通路激活剂Li Cl处理T24细胞,MTT法检测细胞增殖能力,Western blot检测GSK3β(try216)磷酸化水平及β-catenin、c-Myc和cyclin D1蛋白水平,克隆形成试验检测细胞的克隆形成能力。结果显示,过表达Hepa CAM后,能够抑制T24细胞的生长,下调β-catenin、c-Myc及cyclin D1的m RNA和蛋白的表达水平。5,10,20μmol/L的Li Cl作用细胞2 h后,均可促进T24细胞的增殖。10μmol/L的Li Cl作用2 h后能够降低GSK3β的磷酸化水平,促进Wnt信号通路的活化。10μmol/L的Li Cl与Ad-GFP-Hepa CAM联合处理细胞后,能够逆转Hepa CAM对β-catenin、c-Myc及cyclin D1蛋白水平和细胞增殖的抑制作用。该研究表明,Hepa CAM可通过Wnt/β-catenin信号通路抑制膀胱癌细胞T24的增殖。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, re- spectively. 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 LiC1. MTT was used to detect cell growth, the phosphorylation level of GSK3β（try216） and the protein level of D-catenin, c-Myc and cy- clinD1 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 LiC1 for 2 h, cells proliferation was promoted and 10 μmol/L of LiC1 could decrease the phosphoryla- tion level of GSK3β, leading to activation of Wnt/β-catenin pathway. Moreover, combination treatment of 10μmol/L of LiC1 and Ad-GFP-HepaCAM could reverse the inhibition effect of HepaCAM on protein expression levels of β-catenin, c-Myc and cyclinD 1 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.

关 键 词：HEPA CAM WNT/Β-CATENIN 膀胱癌细胞 T24 

分 类 号：R737.14[医药卫生—肿瘤]