摘要
目的研究亚砷酸钠抑制肝癌(HCC)细胞增殖是否与早幼粒细胞白血病(PML)蛋白表达有关。方法免疫组织化学法、免疫荧光、蛋白免疫印迹法(Western blot)和荧光定量PCR检测HCC组织PML蛋白及基因表达。结果免疫组织化学分析显示随机选择15例高分化、中度分化和低分化HCC组织和细胞均不同程度表达PML蛋白。Western blot分析发现24例HCC组织、HuH7、HepG2、Hep3B、SMMC-7721和L02细胞均表达PML蛋白。免疫荧光提示HuH7、HepG2、Hep3B、SMMC-7721细胞核均有PML蛋白颗粒,其中HuH7和Hep3B细胞表达PML蛋白较HepG2、SMMC-7721细胞多。24例HCC组织,Hep3B、HepG2、SMCC-7721和HuH7细胞都表达PML基因。亚砷酸钠不仅可下调HuH7和原代HCC细胞表达PML蛋白,而且亚砷酸钠抑制HuH7,HepG2,Hep3B和SMMC-7721细胞生长,随着暴露时间延长亚砷酸钠对HCC细胞抑制作用增强。结论HCC组织和细胞系普遍表达PML基因和蛋白,PML蛋白可能是砷剂直接靶向作用的分子基础。
Objective To investigate whether the sodium arsenite inhibiting proliferation of hepatocellar carcinoma（HCC）cells and having a relation with the expression of promyelocytic leukemia（PML）protein.Methods The immunohistochemistry,Western blot analysis,immunofluorescence assay and quantitative PCR were used to examine the PML gene and protein expression in HCC tissue and cells.Results The immunohis to chemical staining showed that the PML protein was expressed in nucleus of well-differentiated,moderately differentiated and poorly differentiated HCC tissues randomly selected from 15 cases of HCC undergoing hepatic resection.Western blot analysis showed that PML protein was expressed at varying levels in all 24 HCC tissue samples,HuH7,HepG2,Hep3 B,SMMC-7721 and L02cells.The immunofluorescence assay demonstrated that PML protein grains were found in the nucleis of the HuH7,HepG2,Hep3 Band SMCC-7721 cells,in which HuH7 and Hep3Bexpressed more PML proteins than HepG2 and SMMC-7721 cells.24 HCC tissue samples,Hep3 B,HepG2,SMCC-7721 and HuH7cells all expressed PML gene.Sodium arsenite not only down-regulated the PML protein expressed in HuH7 and primary HCC cells,but also inhibited the HCC cell growth of HepG2,Hep3 B,HuH7and SMMC-7721,with the exposure time extension,the inhibiting effect of sodium arsenite on HCC cells was enhanced.Conclusion Both HCC cell lines and tissues generally express ther PML protein and gene,PML protein may serve as the molecular basis for the direct targeting effects of arsenical agents.
出处
《重庆医学》
CAS
北大核心
2016年第12期1591-1594,共4页
Chongqing medicine
基金
重庆市卫生和计划生育委员会局会中医药科技项目资助(ZY20132047)
