真核表达载体介导白细胞介素24在HepG2细胞的表达及其体外抗肿瘤效应研究  

Expression of interleukin （IL）-24 in hepatocellular carcinoma cell line HepG2 mediated by eukaryotic vector and the anti-tumor effect of IL-24 in vitro

作　　者：于培霞[1] 杨云[1] 王桂琴[1] 

机构地区：[1]山西医学科学院山西大医院检验科,太原030032

出　　处：《国际生物制品学杂志》2014年第1期10-14,共5页International Journal of Biologicals

摘　　要：目的 研究真核表达载体pcDNA3.1（＋）介导白细胞介素（interleukin,IL） 24在肝癌细胞系HepG2细胞中表达的可行性,以及IL-24体外对HepG2细胞的抗瘤效应和可能机制.方法 采用脂质体转染法将重组质粒pcDNA3.1（＋）-IL-24及空质粒分组转染HepG2细胞,在倒置显微镜下观察细胞形态变化.用噻唑蓝比色方法测量细胞增殖指数,流式细胞仪研究细胞早期凋亡率及其细胞周期分布.结果 IL-24 mRNA成功表达于HepG2细胞中.重组质粒转染组可见明显的凋亡细胞形态,48 h增殖抑制率（F=27.058,P＜0.01）、72 h增殖抑制率（F =63.481,P ＜0.01）和早期细胞凋亡率（F=326.220,P ＜0.01）与对照组的差异均有统计学意义.细胞分布呈明显的G2/M周期阻滞.结论 真核表达载体pcDNA3.1（＋）可以有效地介导IL-24在HepG2细胞的表达.IL-24对HepG2细胞有明显的增殖阻滞和凋亡诱导效应,G2/M细胞周期阻滞或许是IL-24抗瘤效应的机制.Objective To study the feasibility of a eukaryotic vector mediating expression of MDA-7/IL-24 in hepatocellular carcinoma cell line HepG2,the anti-tumor effect of MDA-7/IL-24 in HepG2 cells and possible working mechanism for the tumor-suppressor activity.Methods The recombinant plasmid pcDNA3.1 （＋）-IL-24 and empty plasmid pcDNA 3.1 （＋） were transfected into HepG2 cells by liposome transfection,respectively.Morphological changes of apoptosis were observed under inverted microscope.The proliferation-inhibiting effect of IL-24 in HepG2 cells was observed with Thiazolyl blue assay.Apoptosis ratio and cell-cycle were analyzed by flow cytometry.Results The mRNA of IL-24 was detected in HepG2 cells transfected with pcDNA3.1 （＋）-IL-24 successfully.The typically morphological changes of apoptotis of cells transfected with the target gene were observed obviously.The proliferation-inhibiting rates at 48 h posttransfection （F =27.058,P ＜ 0.01）,and 72 h post-transfection （F =63.481,P ＜ 0.01）,and apoptosis indexes （F =326.220,P ＜ 0.01） in IL-24 group were significantly higher than those in control groups,with the proportion of cells in the phase of G2/M in IL-24 group being higher,too.Conclusions The recombinant eukaryotic vector pcDNA3.1 （＋） can mediate expression of MDA-7/IL-24 in HepG2 cells effectively.IL-24 displays growth-inhibiting and apoptosis-inducing activity in HepG2 cells and cell circle arrest in the phase of G2/M may be the working mechanism of the anti-tumor effect.

关 键 词：白细胞介素类 HEPG2细胞 基因表达 细胞周期 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]