摘要
目的:探讨转化生长因子β1(transforming growth factorβ1,TGF-β1)体外对肝癌细胞干性特征的影响。方法:以转化H-ras的鼠肝肿瘤细胞LPC-H-ras和人肝癌细胞株PLC/PRF/5为研究对象,利用MTT法和多细胞球体培养法检测TGF-β1对肿瘤起始细胞增殖的抑制作用,FCM法检测细胞表面分子CD133的表达以及活性氧(reactive oxygen species,ROS)的水平;实时荧光定量PCR(real-time fluorogentic quantitative-PCR,RFQ-PCR)检测干细胞样基因的表达水平。结果:TGF-β1在体外能够改变LPC-H-ras细胞的形态并抑制细胞的生长;与未处理组相比,TGF-β1作用后,LPC-H-ras细胞表面CD133的表达降低,多细胞球体形成能力减弱,干细胞样基因的表达受到抑制;同样,在PLC/PRF/5细胞中,TGF-β1也能抑制CD133的表达、AFP基因表达以及多细胞球体的形成;另外,在LPC-H-ras细胞中,TGF-β1可诱导ROS表达升高,ROS的抑制剂N-乙酰半胱氨酸具有削弱TGF-β1诱导CD133表达下调的作用。结论:TGF-β1能够抑制肝癌起始细胞的增殖及干性,其作用机制可能与ROS表达上调有关。
Objective： To investigate the effect of transforming growth factor β1 （TGF-β1） on the stem-like phenotype of liver tumor cells in vitro. Methods： The inhibitory effects of TGF-β1 on the proliferation of liver tumor-initiating cells—mouse liver tumor cell LPC-H-ras and human liver tumor cell PLC/PRF/5 were evaluated by using MTT method and spheroid formation analysis. The expression of surface marker CD133 and reactive oxygen species （ROS） was detected by flow cytometry （FCM）. The relative levels of stem-cell like genes were detected by real-time fluorogenic quantitative-PCR （RFQ-PCR）. Results： TGF-β1 changed the morphology and inhibited the growth of LPC-H-ras cells in vitro. Compared with the untreated cells, the expression of CD133 in LPC-H-ras cells and their ability of spheroid formation induced by TGF-β1 were significantly decreased. In addition, the expressions of stem-cell like genes were inhibited after treatment with TGF-β1 in these cells. Similarly, the expression levels of CD133 and AFP mRNA and the spheroid formation ability were all down-regulated in PLC/PRF/5 cells treated with TGF-β1. Furthermore, the ROS level in LPC-H-ras cells mediated by TGF-β1 was increased, but it was decreased by using antioxidant N-acetylcysteine. Conclusion： TGF-β1 inhibits the proliferation and stem-like properties of mouse liver tumor-initiating cells by up-regulating ROS production.
出处
《肿瘤》
CAS
CSCD
北大核心
2012年第4期227-233,共7页
Tumor
关键词
肝肿瘤
实验性
肿瘤干细胞
转化生长因子Β1
CD133
活性氧
Liver neoplasms
experimental
Neoplatic stem cell
Transforming growth facter beta 1
CD133
Reactive oxgen species
