摘要
目的 :检测酸性亮氨酸核磷酸蛋白32B(acidic leucine-rich nuclear phosphoprotein 32B,ANP32B)在肝细胞肝癌组织中的表达水平,并探讨其在肝癌细胞生长中的作用。方法:利用国际权威公共数据库,分析肝细胞肝癌组织中ANP32B mRNA的表达及其与患者生存期间的关系;用蛋白免疫印迹法检测患者肝癌组织样本和敲除ANP32B的肝癌细胞系中的ANP32B蛋白表达水平;并用RNA干扰技术在肝癌细胞系中沉默ANP32B的表达,通过计数检测细胞生长及克隆形成,用流式细胞仪检测其细胞周期和细胞凋亡。结果:通过数据库及肝癌组织样本分析发现,肝癌组织中的ANP32B mRNA表达值(9.763±0.04)相较于癌旁组织(9.293±0.03)明显升高,蛋白表达升高1.78倍,且ANP32B高表达的肝癌患者与ANP32B低表达的患者相比,5年总生存率明显下降,差异有统计学意义(P<0.01)。体外实验发现,2组敲除ANP32B的肝癌细胞和对照组肝癌细胞相比,细胞生长速度明显减慢(29.9×10~5比23.2×10~5,15.5×10~5),克隆形成数目减少(300个比146个比135个),细胞周期发生S期阻滞(47.9%比61.5%比57.1%),细胞凋亡数略有增加(1.8%比7.3%比4.9%)。结论:ANP32B在肝癌组织中高表达,在肝癌细胞系中沉默ANP32B表达后可明显抑制细胞生长,细胞周期发生阻滞,细胞凋亡增加。
Objective: To investigate the ANP32 B(acidic leucine-rich nuclear phosphoprotein 32 B)expression in hepatocellular carcinoma and determine its clinical significance. Methods: The mRNA expression of ANP32 B in hepatocellular carcinoma and the overall survival were analyzed using international public datasets. The protein expression of ANP32 B in hepatocellular carcinoma tissue was detected by immunoblotting. ANP32 B was knockdown by shRNA in HCC cell lines, and cell growth and colony formation were analyzed;cell cycle and apoptosis were detected by flow cytometry. Results: ANP32 B mRNA and protein level were highly expressed in hepatocellular carcinoma, and the higher ANP32 B mRNA level was asso-ciated with shorter survival. Knockdown of ANP32 B in HCC cell lines inhibited cell growth and colony formation, induced cell cycle arrest and cell apoptosis. Conclusions: ANP32 B is highly expressed in hepatocellular carcinoma. Knockdown of ANP32 B in HCC cell lines inhibited cell growth and colony formation, induced cell cycle arrest and cell apoptosis.
作者
朱迪
朱晓娜
杨烁
何平
沈少明
余韵
ZHU Di;ZHU Xiaona;YANG Shuo;HE Ping;SHEN Shaoming;YU Yun(Center of Molecular Medicine,Ruijin Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China;Department of Hematology,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200127,China)
出处
《诊断学理论与实践》
2019年第2期170-176,共7页
Journal of Diagnostics Concepts & Practice
基金
国家自然科学基金(81772936)
关键词
酸性亮氨酸核磷酸蛋白32B
肝癌
细胞生长
Acidic leucine-rich nuclear phosphoprotein 32B(ANP32B)
Hepatocellular carcinoma
Cell growth
