抗肺癌小分子抑制剂的分子机制研究

发布时间：2018-11-11 16:01

【摘要】：肺癌是一种恶性程度很高,诊断与治疗都很困难的呼吸道肿瘤。据报道近50年来的发病与死亡率均显著升高,男性肺癌发病及死亡率均占据所有恶性肿瘤的第一位,在女性中的发病与死亡率占第二位,严重危害着人们的生命与健康。寻找新型肿瘤抑制剂一直是重要研究课题。人类肺癌25%以上是由ras基因突变引起的,针对Ras蛋白的靶向治疗已经成为了寻找抗肺癌新药的研究热点。本研究采用细胞学和分子生物学技术观察针对Ras蛋白设计的小分子抑制剂NSC290956,1-Thia-4,8-diazaspiro[4.5]decan-3-1,8-[3-(2-chloro-10Hphenothiazin-10-yl)propyl]-hydrochlo-ride,在k-ras突变肺癌细胞系NCI-H358中通过体外实验研究凋亡,自噬和周期调控等,探究其对肿瘤细胞的抑制作用和相关作用机制。本文的主要结论是:NSC290956体外对K-ras突变肺癌NCI-H358细胞的增殖具有明显的抑制作用,NSC290956可能通过增加细胞内ROS,上调Bax蛋白表达,降低细胞线粒体膜电位,促使线粒体细胞色素C释放入胞浆,继而启动Caspase级联反应,诱导A549和H358细胞凋亡,最终抑制肺癌细胞的增殖。NSC290956通过抑制A549和H358细胞株的Ras/Raf/MEK/ERK信号通路从而抑制癌细胞的发生、发展。通过流式细胞术和western blot证明NCS290956诱导细胞凋亡与细胞周期阻滞有关。NSC290956通过诱导H358细胞株的自噬作用从而抑制癌细胞的生长与增殖。
[Abstract]:Lung cancer is a highly malignant, difficult diagnosis and treatment of respiratory tract tumors. It has been reported that the incidence and mortality of lung cancer have increased significantly in the past 50 years. The morbidity and mortality rate of lung cancer in men are the first among all malignant tumors, and the morbidity and mortality rate in women is the second, which seriously endangers people's lives and health. Searching for new tumor inhibitors has been an important research topic. More than 25% of human lung cancer is caused by mutation of ras gene. Targeted therapy for Ras protein has become a hot spot in search of new anti-lung cancer drugs. In this study, cytological and molecular biological techniques were used to observe the design of NSC290956,1-Thia-4,8-diazaspiro [4.5] decan-3-1,8- [3- (2-chloro-10Hphenothiazin-10-yl) propyl] -hydrochlo-ride, for Ras protein. Apoptosis, autophagy and cycle regulation were studied in k-ras mutant lung cancer cell line NCI-H358 in vitro. The main conclusions of this study are as follows: NSC290956 can inhibit the proliferation of K-ras mutant lung cancer NCI-H358 cells in vitro, and NSC290956 may increase the expression of Bax protein and decrease the mitochondrial membrane potential by increasing the expression of ROS, in the cells. Mitochondrial cytochrome C was released into the cytoplasm, and then Caspase cascade reaction was initiated, which induced apoptosis of A549 and H358 cells. NSC290956 inhibits the development of cancer cells by inhibiting the Ras/Raf/MEK/ERK signaling pathway of A549 and H358 cells. Flow cytometry and western blot showed that apoptosis induced by NCS290956 was related to cell cycle arrest. NSC290956 inhibited the growth and proliferation of H358 cells by inducing autophagy.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R979.1

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