摘要
目的 观察ZBP-89对八聚体结合转录因子4(Oct4)信号通路和肝癌细胞干性的影响。方法 采用免疫组织化学染色法检测120例肝癌组织中ZBP-89、CD133和上皮细胞黏附因子(EpCAM)的表达,分析ZBP-89的表达水平与CD133、EpCAM的相关性。通过ZBP-89过表达的慢病毒载体和小干扰RNA(siRNA)调节Hep3B细胞中ZBP-89的表达,实时定量聚合酶链反应(Real-time PCR)和Western blot检测Oct4、CD133和EpCAM的表达。结果 在肝癌组织中ZBP-89的表达水平和CD133、EpCAM的表达呈负相关(χ2=45.000,P<0.05;χ2=26.020,P<0.05)。当Hep3B细胞中的ZBP-89表达升高时,Oct4、CD133和EpCAM的表达下降,其中mRNA水平分别为对照组的(0.47±0.03)、(0.63±0.01)、(0.53±0.02)倍(P均<0.05),当ZBP-89的表达下降时Oct4、CD133和EpCAM的表达升高,其中mRNA水平分别为对照组的(2.04±0.03)、(1.62±0.03)、(1.79±0.05)倍(P均<0.05)。Western blot与Real-time PCR结果相一致。结论 ZBP-89可通过抑制Oct4信号通路抑制肝癌细胞的干性。
Objective To investigate the effect of zinc-finger binding protein-89 (ZBP-89) on octamer binding transcription factor 4 (Oct4) signaling pathway and liver cancer stemness.Methods Immunohistochemical staining was used to detect the expression of ZBP-89, CD133 and epithelial cell adhesion molecule (EpCAM) in 120 cases of hepatocellular carcinoma (HCC), and the correlation between ZBP-89 expression and CD133 and EpCAM was analyzed. The expression of ZBP-89 in Hep3B cells was regulated by ZBP-89 overexpressing lentiviral vector and small interfering RNA (siRNA), and the expression of Oct4, CD133 and EpCAM was detected by real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting.Results The expression of ZBP-89 was negatively correlated with the expression of CD133 and EpCAM in HCC tissues (χ2=45.000, P<0.05;χ2=26.020, P<0.05). When the expression of ZBP-89 in Hep3B cells was increased, the expression of Oct4, CD133 and EpCAM decreased, and the mRNA levels of Oct4, CD133 and EpCAM were (0.47±0.03), (0.63±0.01) and (0.53±0.02) times of the control group respectively (all P<0.05). When the expression of ZBP-89 decreased, the expression of Oct4, CD133 and EpCAM increased, and the mRNA levels of Oct4, CD133 and EpCAM were (2.04±0.03), (1.62±0.03) and (1.79±0.05) times of the control group respectively (all P<0.05). The results of Western blotting were consistent with the results of Real-time PCR.Conclusion ZBP-89 can inhibit the stemness of liver cancer cells by inhibiting the Oct4 signaling pathway.
作者
刘利平
何前进
吴宗泽
韦超盈
吴红
鲍世韵
Liu Liping;He Qianjin;Wu Zongze;Wei Chaoying;Wu Hong;Bao Shiyun(Department of Hepatobiliary and Pancreas Surgery, the Second Clinical Medical College of Jinan University (Shenzhen People’s Hospital), Shenzhen 518020, China;Department of General Surgery, the Center Hospital of Huanggang, Huanggang 438000, China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2019年第2期315-318,共4页
Chinese Journal of Experimental Surgery
基金
广东省自然科学基金 (2017A030313846)
深圳市科技创新基础研究项目 (JCYJ20160422152408705).
关键词
ZBP-89
八聚体结合转录因子4
肝癌
上皮细胞黏附因子
CD133
Zinc-finger binding protein-89
Octamer binding transcription factor 4
Hepatocellular carcinoma
Epithelial cell adhesion molecule
CD133
