Notch 1信号通路对人肝癌SMMC 7721细胞中CD133^+细胞增殖和凋亡的影响  

Role of Notch 1 signaling in the proliferation and Die depth of CD133^+ cells from human hepatoma SMMC 7721 cells

作　　者：吴乐乐[1] 俞广进[1] 陈双升 熊奇如[1] 

机构地区：[1]安徽医科大学第一附属医院普通外科,合肥230022

出　　处：《安徽医科大学学报》2014年第12期1701-1705,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省科技厅年度重点科研项目(编号:12070403066)

摘　　要：目的研究人肝癌SMMC 7721细胞中CD133+细胞的干细胞特性,初步探讨调控Notch 1信号通路对CD133+细胞亚群增殖和凋亡的影响。方法流式分选和检测SMMC 7721细胞中的CD133+细胞。利用软琼脂集落实验及裸鼠成瘤实验验证CD133+细胞的干细胞特性。RT-PCR法检测CD133+细胞中Notch信号系统的表达;将分选出来的CD133+细胞分为激活组(加入Notch 1激活剂Jagged 1)、抑制组(加入Notch 1抑制剂DAPT)和空白对照组(加入PBS缓冲液),RT-PCR法检测3组细胞Hes-1基因的表达,MTT法和流式细胞仪检测各组细胞增殖能力和凋亡能力。结果成功分选出CD133+亚群细胞;软琼脂集落实验和裸鼠成瘤实验表明CD133+细胞较CD133-细胞具有较强集落形成能力和体内成瘤能力;RT-PCR检测CD133+细胞中仅表达Notch 1/Jagged 1信号通路;激活组Hes-1表达强度增强,抑制组表达强度减弱,差异有统计学意义(P<0.05)。MTT检测激活组、抑制组和空白对照组吸光度值差异均有统计学意义(P<0.05)。流式细胞仪检测激活组总凋亡率大于空白对照组,反之,抑制组总凋亡率小于空白对照组,各组差异有统计学意义(P<0.05)。结论 CD133+细胞是人肝癌SMMC 7721细胞中干细胞标志物之一,Notch 1信号通路对CD133+细胞的增殖和凋亡发挥着重要作用。Objective To study the stem cell properties of CD133 ＋ cells from the human hepatoma SMMC 7721 cells and investigate the effect of Notch 1 signaling pathway on the proliferation and apoptosis of CD133 ＋ cells.Methods The CD133 ＋ cells from SMMC 7721 cells were sorted out and detected by flow cytometer.Soft agar colony formation and tumor formation in nude mice were used to validate the stem cell properties of CD133 ＋ cells.The Notch signaling system of CD133 ＋ cells was measured by RT-PCR.The Jagged 1,an activator of Notch signaling,and the γ-secretase inhibitor （DAPT） were added into the mediums of the CD133 ＋ cells respectively,the control was added with PBS buffer.Then the level of each Hes-1 mRNA was measured by RT-PCR,MTT assay and Flow cytometry were used to detect the proliferation and apoptosis of CD133 ＋ cells.Results The CD133 ＋ cells were sorted out from the SMMC 7721 cells successfully.Compared to CD133-cells,CD133 ＋ cells had stronger abilities of colony formation in vitro and tumorigenic in vivo.Only the Notch 1/Jagged 1 expressed in the CD133 ＋ cells.Expression of Hes-1 mRNA in activator group and inhibitor group had significant difference compared with control group（P ＜0.05）,A value of CD133 ＋ cells cultured in activator group and inhibitor group had significant difference compared with control group （P ＜ 0.05）,and the total apoptosis rates in activator group and inhibitor group had significant difference compared with control group （P ＜ 0.05）.Conclusion The CD133 ＋ cells have the stem cell properties and the CD133 ＋ is a label of the stem cell in human SMMC 7721 cells.The Notch 1 signaling pathway plays an important role in the proliferation and apoptosis of the CD133＋ cells.

关 键 词：CD133+ NOTCH 1信号通路 肝癌 

分 类 号：R735.7[医药卫生—肿瘤]