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目的:研究胰岛素样生长因子Ⅰ型受体(insulin-like growth factor-Ⅰ receptor,IGF-ⅠR)反义基因对SMMC-7721肝癌细胞株形态学的影响,为肝癌的发病机制提供理论基础。 方法:构建IGF-ⅠR正、反义基因真核表达载体,脂质体介导将其转入SMMC-7721肝癌细胞株,HE染色、电镜检查肝癌细胞株形态学的变化,流式细胞仪检测细胞周期、凋亡。 结果:HE染色IGF-ⅠR反义细胞与正义细胞、7721细胞无明显变化。扫描电镜:IGF-ⅠR正义细胞异形性明显,微绒毛较反义细胞、7721细胞显著,透射电镜:7721细胞大小不等,细胞器增多,核浆比例增大。IGF-ⅠR反义细胞胞质内可见微腺腔、灶性坏死、髓鞘样结构。流式细胞仪检测IGF-ⅠR反义细胞G1期细胞明显增加(0.64),S期细胞减少(0.19),而且凋亡增加(0.06)。IGF-ⅠR正义细胞S期细胞增加(0.35),凋亡减少(0.01)。反义细胞与正义细胞、7721细胞比较有非常显著性差异(P<0.01)。 结论:反义细胞中出现的异常形态学改变可能与IGF-ⅠR反义基因抑制细胞增生、促进细胞凋亡有密切关系。
Objective: To study the effect of insulin-like growth factor-1 receptor (IGF-ⅠR) antisense gene on the morphology of SMMC-7721 hepatocellular carcinoma cell line and to provide a theoretical basis for the pathogenesis of hepatocellular carcinoma. Methods: Eukaryotic expression vector of sense and antisense IGF-ⅠR gene was constructed and transfected into SMMC-7721 hepatoma cell line by lipofectamine. HE staining and electron microscopy were used to examine the morphological changes of hepatocellular carcinoma cell line. Flow cytometry Cell cycle, apoptosis. Results: HE staining of IGF-ⅠR antisense and sense cells, 7721 cells without significant change. Scanning electron microscopy: IGF-ⅠR positive cells were significantly morphological heterogeneity, microvilli antisense cells, 7721 cells significantly, transmission electron microscopy: 7721 cells ranging in size, organelles increased, the proportion of nuclear plasma increased. IGF-Ⅰ R antisense cell cytoplasm can be seen within the micro-gland cavity, focal necrosis, myelin-like structure. Flow cytometry showed that the number of G1 phase cells increased significantly (0.64), the number of S phase cells decreased (0.19), and the apoptosis increased (0.06) in IGF-ⅠR antisense cells. S-phase cells in IGF-IR positive cells increased (0.35) and apoptosis decreased (0.01). There was a significant difference between antisense and sense cells and 7721 cells (P <0.01). Conclusion: The abnormal morphological changes in antisense cells may be closely related to the inhibition of cell proliferation and cell apoptosis by IGF-ⅠR antisense gene.