稳定干扰SET肝细胞株的生物学鉴定

Biologic identification of SET protein-deficient L-02 cells

目的:对稳定干扰SET(patient SE translocation)的肝细胞株进行生物学性状鉴定,为研究SET蛋白在肝细胞中的作用奠定基础。方法:培养人肝L-02细胞、慢病毒载体介导的稳定干扰(siRNA)SET L-02肝细胞及psiRNAc L-02肝细胞(空载体转染细胞),于倒置显微镜下观察细胞形态,利用MTT吸光度与细胞数目间关系绘制生长曲线,采用染色体畸变试验观察染色体有无结构异常,软琼脂克隆实验检测细胞的恶性转化。结果:与正常肝细胞比较,稳定干扰SET肝细胞及psiRNAc肝细胞的生长形态无明显变化,3组细胞的生长曲线趋于一致。染色体畸变分析表明SET基因沉默的人肝L-02细胞染色体结构与数目未发生明显改变。软琼脂克隆实验表明SET基因沉默的人肝L-02细胞未发生恶性转化。结论:SET siRNA的导入未引起细胞生物学特征的改变,遗传性状保持稳定,说明所构建的SE7基因沉默的人肝L-02细胞模型是稳定的,为后续的研究提供了工具细胞。

OBJECTIVE：This study aimed to identify the biological characteristics of SET protein-deficient L-02 cells which could lay a foundation for the role of SET protein in normal L-02 cells.METHODS：Normal L-02 cells,SET protein-deficient L-02 cells（stably expressing SET siRNA）,siRNAc transfected L-02 cell were cultured and the growth and the morphology of these three groups of cells were examined and analyzed.The growth curves of the cells were generated on the data from MTT assay.Chromosome structure was detected by chromosome aberration analysis. Malignant transformation was identified by soft agar cloning test.RESULTS：Compared with normal L-02 cells,no obvious changes of cytomorphology and growth curves were observed for SET protein-deficient L-02 cells and siRNAc transfected L-02 cells.Chromosome aberration analysis revealed that transfection of SET siRNA did not cause obvious changes in the structure and numbers of chromosomes in L-02 cells.Soft agar cloning test showed that transfection of SET siRNA did not cause malignant transformation of the cells.CONCLUSION：Transfection of SET siRNA did not significantly alter the biologic characteristics of the cells,indicating that the established SET-deficient cells can be used as a model for further studies.