干扰素-γ基因转导前后人肝癌细胞HepG1表型变化及基因表达图谱差异的研究

Study on the Difference of Genotype and Gene Expression Profile of Human Hepatocellular Carcinoma Cell HepG1 Transduced with Interferon-r Gene

目的：比较人肝癌细胞经干扰素（ＩＦＮ）－γ基因转导前后细胞表型及基因表达图谱的差异。方法：①用扫描电镜、流式细胞仪等方法观察ＩＦＮ－γ基因转导前后人肝癌细胞ＨｅｐＧ１表型的变化；③应用微阵列膜片分别与未转导及转导了ＩＦＮ－γ基因的ＨｅｐＧ１ ｍＲＮＡ进行杂交，同时设一转导空白载体ＬＸＳＮ的ＨｅｐＧ１ ｍＲＮＡ与膜片杂交作为对照，比较杂交信号，获得表达差异的基因。结果：①ＩＦＮ－γ基因转导的ＨｅｐＧ１细胞生长速度减慢，表面绒毛结构数量增多，绒毛变细、变长，ＨＬＡⅠ类和Ⅱ类分子表达增高；②ＩＦＮ－γ基因转导后，ＨｅｐＧ１细胞有多种基因的表达发生变化，其中一些基因较未转导ＩＦＮ－γ时表达增高，一些较未转导ＩＦＮ－γ时表达减低。表达发生变化的基因主要为与细胞周期和细胞凋亡相关的基因。结论：ＩＦＮ－γ基因修饰ＨｅｐＧ１细胞后使细胞表型发生变化，并且在基因表达上也发生改变。运用微阵列技术为进一步了解这些基因的相互关系及其与肿瘤生物学行为的关系奠定了基础，也可为优化肿瘤免疫基因治疗提供线索。

Background/Aims: To compare the difference of phenotype and gene expression profile of human hepatocellular carcinoma cells transduced or not with interferon (IFN)-γ gene. Methods: 1. Using scanning microscope and FACS to detect the change of phenotype of HepG1 transduced with IFN-γ gene; 2. Using microarray membranes to hybridize with HepG1, HepG1-IFN-γ, and HepG1-LXSN mRNA, respectively, then compared their hybridized signal and the difference of their gene expression. Results: 1. The growth speed of HepG1-IFN-γ decreased, the amount of surface microvilli increased and surface microvilli became thinner and elongated. the expression level of HLA I and II molecules also increased; 2. After transduced with IFN-γ gene, the expression level of many genes in HepG1 were changed, some decreased and others increased. The changed genes were mainly associated with cell cycle and apoptosis. Conclusions: HepG1 transduced with IFN-γ gene could produce changes of phenotype and gene expression. Using microarrays technology enables us to learn more information about the interrelationship among those changed genes and their association with tumorigenicity, it also provides us some clues to the advantageous tumor immuno-gene therapy strategy.