抗Cyclin D1胞内抗体与p16联合抗肿瘤效应研究

发布时间：2018-06-20 12:26

本文选题：Cyclin + D1　；参考：《吉林大学》2008年硕士论文

【摘要】： 细胞周期蛋白Cyclin D1和周期蛋白依赖性蛋白激酶抑制剂p16分别是细胞周期G1/S转换的正负调节因子。在正常情况下,Cyclin D1和p16保持平衡,但在肿瘤细胞中,却存在Cyclin D1的过量表达和p16的广泛失活。由于Cyclin D1的过量表达和p16的失活在肿瘤细胞中同时存在,而且它们的下游分子都是CDK4/6,所以同时抑制Cyclin D1的活性和恢复p16的功能,可以使对CDK4/6的抑制作用得到叠加,因此开展它们的联合治疗可能是一个可行的基因治疗策略。胞内抗体技术是基于抗体工程和胞内信号传导而衍生出来一项新的实现细胞内重要靶蛋白表型敲除的技术。在实验中,我们将前期构建的靶向Cyclin D1的内质网滞留型的胞内抗体基因和野生型p16基因分别单独和联合导入Cyclin D1过量表达和p16纯合缺失的MCF-7细胞中。实验发现,这些外源基因在细胞中都得到表达,并且表达的胞内抗体能与细胞内的Cyclin D1结合。同时发现,它们的转染都能引起细胞G1期的阻滞,诱导细胞凋亡和抑制细胞增殖,而且联合转染比单独转染效果更显著。这为基因的联合治疗开辟了新的途径,具有一定的应用价值。
[Abstract]:Cyclin D1 and cyclin dependent protein kinase inhibitor p16 are positive and negative regulators of cell cycle G1 / S transition, respectively. Under normal conditions, Cyclin D1 and p16 remain in balance, but in tumor cells, there is overexpression of Cyclin D1 and widespread inactivation of p16. Because the overexpression of Cyclin D1 and the inactivation of p16 exist simultaneously in tumor cells and their downstream molecules are CDK4 / 6, the inhibition of the activity of Cyclin D1 and the restoration of p16 function at the same time can result in the superposition of the inhibitory effect on CDK4 / 6. Therefore, the development of their combined therapy may be a feasible gene therapy strategy. Intracellular antibody technology is a new technique based on antibody engineering and intracellular signal transduction to realize the knockout of important target protein phenotypes in cells. In the experiment, the previously constructed endoplasmic reticulum (ER) retention type antibody gene and wild-type p16 gene were introduced into MCF-7 cells with overexpression of Cyclin D1 and homozygous deletion of p16, respectively. It was found that these exogenous genes were expressed in cells, and the expressed intracellular antibodies could bind to Cyclin D1. It was also found that their transfection could induce G1 phase arrest, induce apoptosis and inhibit cell proliferation, and the effect of co-transfection was more significant than that of single transfection. This opens up a new way for gene combination therapy and has certain application value.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392;R730.5

【参考文献】