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探讨肝细胞粘附分子(hepatocyte cell adhesion molecule,hepa CAM)对膀胱癌细胞BIU-87增殖活力的影响以及其调节机制。将BIU-87细胞分为对照组、空载组、hepa CAM组、LY294002组及hepa CAM与LY294002联用组,噻唑蓝法(methyl thiazolyl tetrazolium,MTT)检测细胞的增殖抑制率;Real-time PCR检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达;Western blot法检测p-FOXO1的表达;同时用LY294002处理BIU-87细胞后Western Blot检测p-FOXO1、PCNA的表达。结果显示,单独运用hepa CAM过表达腺病毒或LY294002均能够明显抑制膀胱癌BIU-87细胞的增殖活力,hepa CAM过表达腺病毒与LY294002联用更加显著的增强了单独作用的抑制效果(p<0.05);Western Blot显示,hepa CAM基因的过表达使p-FOXO1的表达量显著降低(p=0.001);hepa CAM过表达腺病毒与LY294002联用后,与hepa CAM或LY294002分别作用相比,p-FOXO1(p=0.014,p=0.047)和PCNA(p=0.002,p=0.004)的表达量降低明显;Realtime PCR结果显示,LY294002处理BIU-87细胞后,PCNA的表达明显减低(p=0.003),加入hepa CAM过表达腺病毒后更为显著地增强LY294002对PCNA的抑制作用(p=0.001)。本课题得出结论,hepa CAM基因过表达后通过下调p-FOXO1从而抑制膀胱癌BIU-87细胞的增殖活力,此作用与PCNA的表达量下调有关。
Abstract：This study aimed to explore the effect of hepa CAM(hepatocyte cell adhesion molecule) on cell proliferation in bladder cancer cell BIU-87 and the potential mechanism. We divided BIU-87 cells into control group,no-load group, hepa CAM group, LY294002 group, and hepa CAM+LY294002 group, then we detected the inhibition rate of BIU-87 cell by MTT(methyl thiazolyl tetrazolium) assay, identified the gene expression of PCNA(proliferating cell nuclear antigen) by Real-time PCR, determine the protein expression of p-FOXO1 by Western blot, and measured the expression of p-FOXO1 and PCNA after treatment with LY294002 by Western Blot. The results showed that proliferation of bladder cancer BIU-87 cell was remarkably decreased by the adenovirus of over-expressed hepa CAM or LY294002 treatment alone, and combined treatment with adenovirus of hepa CAM and LY294002 decreased the cell viability more significantly(p<0.05). Western blot assay showed that over-expression of hepa CAM reduced the protein expression of p-FOXO1(p=0.001) markedly, but compared with the respective of hepa CAM or LY294002, the expression decreased significantly(p-FOXO1: p=0.014, p=0.047; PCNA: p=0.002, p=0.004) after the use of combined injection by ad-hepa CAM and LY294002. Real-time PCR showed that the expression of PCNA decreased significantly(p=0.003) after the treatment of BIU-87 cell by LY294002, while the inhibition effect of LY294002 on PCNA got stronger after over-expressed hepa CAM virus was added(p=0.001). Our study showed that the over-expressed hepa CAM inhibited the proliferation of BIU-87 cell through downregulating p-FOXO1,which may be related to the decreased expression of PCNA.
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基本信息:
DOI：10.13417/j.gab.035.000756
中图分类号:R737.14
引用信息:
[1]唐敏,赵燕,陈娥,等.hepaCAM通过下调p-FOXO1/PCNA调节人膀胱癌BIU-87细胞的增殖活力[J].基因组学与应用生物学,2016,35(04):756-762.DOI:10.13417/j.gab.035.000756.
基金信息:
国家自然科学基金(81072086)项目资助
2016-04-25
2016-04-25