上调神经生长因子-β对人胆管癌细胞QBC939、生物学特性的影响  

Effect of NGF-β up-regulation on the biological characteristics of human cholangiocarcinoma cell line QBC939

作　　者：岳秀敬[1] 徐莉君 张锐[1] 许磊波[1] 刘超[1] 

机构地区：[1]中山大学孙逸仙纪念医院肝胆胰外科,广州510120 [2]安徽省淮南市东方总医院内科,安徽淮南232001

出　　处：《岭南现代临床外科》2012年第5期250-253,共4页Lingnan Modern Clinics in Surgery

基　　金：广东省自然科学基金(915104070100002)

摘　　要：目的探讨神经生长因子-β上调对人胆管癌细胞QBC939增殖、凋亡、细胞周期及克隆形成能力等生物学特性的影响。方法构建稳定转染pEGFP-N1-NGF的QBC939细胞,上调神经生长因子-β的表达(高表达组),转染pEGFP-N1-NC组的QBC939细胞作为对照组。以Western blot方法检测细胞转染效果,细胞增殖实验(CCK-8实验)检测细胞增殖能力,单克隆形成实验检测细胞克隆形成能力,流式细胞术检测细胞周期及细胞凋亡。结果在450nm波长下,24小时测得对照组QBC939细胞的吸光度值为0.409±0.014,高表达组细胞的吸光度值为0.691±0.029;48小时测得对照组QBC939细胞的吸光度值为1.612±0.044,高表达组细胞的吸光度值为2.033±0.005,其差异均具有统计学意义(P<0.05);200倍显微镜下观察,对照组直径大于500um的单细胞克隆数比例为10%,高表达组直径大于500um的单细胞克隆数比例为24%,统计学差异显著(P<0.01);对照组S期细胞数所占比例为(15.643±0.693)%,高表达组S期细胞数所占比例为(40.193±1.671)%,统计学差异显著(P<0.01);对照组细胞早期凋亡率为(15.76±0.97)%,高表达组细胞早期凋亡率为(8.82±0.93)%,具有统计学差异(P<0.05)。结论上调神经生长因子-β能够增强QBC939细胞的增殖与克隆形成能力,能够促进细胞进入S期,抑制细胞凋亡。Objective To investigate the effect of nerve growth factor-β （NGF-β） upregulation in human cholangiocarcinoma cell line QBC939 on its proliferation , apoplosis, cell cycle and cell cloning ability in vitro. Methods QBC939 cells transfeeted with pEGFP-N1-NC were named the control group. QBC939 ceils transfected with the pEGFP-N1-NGF were constructed and it named the over-expression group. The effect of transfection was determined by Western blot method. The proliferative and colony formation ability of QBC939 cells was assessed by CCK-8 assay and sphere fi）rmation assay, respectively. The cell cycle and apoptosis of the QBC939 cells were detected by flow cytometry. Results We measured at 24 hours in the 450urn wavelength, the absorbance value of the control group QBC939 cells was 0.409±0.014, the absorbance value of high expression group was 0.691±0.029. We measured at 48 hours that the absorbance value of the control group QBC939 cells was 1.612±0.044, the absorbance value of the high expression group was 2.033 ±0.005,and the differences were statistically significant （P〈0.05）. We also observed under 200 times magnification, and the proportion of single-cell clones with a diameter greater than 500urn was 10% in the control group, while it was 24% in the high expression group, tile difference was statistically significant （P〈0.01）; the proportion of S phase cells in the control group was （15.643±0.693）%, while it was （40.193±1.671）% in the high expression group, the difference was statistically significant （P〈0.01）. In the control group, the early apoptosis rate was （15.76±0.97）%, while the early apoptosis rate was （8.82±0.93）% in the high expression group, the difference was statistically significant （P〈0.05）. Conclusion Up-regulation of NGF-β can increase the proliferative and colony formation ability of QBC939 cells, promote QBC939 cells to S phase and inhibit cell apoptosis.

关 键 词：神经生长因子-β 细胞增殖 细胞凋亡 细胞周期 克隆形成 转染 

分 类 号：R735.8[医药卫生—肿瘤]