核苷化合物N的抗肿瘤活性及其机制研究
发布时间：2018-12-24 08:17
【摘要】：目的 在本论文中,通过MTT等体外实验观察核苷类化合物N对多个肿瘤细胞系的抗肿瘤活性,以及初步探讨该核苷化合物可能具有的抗肿瘤作用机制,从而为开发新的抗肿瘤化疗药物或发现先导化合物奠定初步的理论基础。 方法 1.通过常规细胞毒性实验MTT法,观察核苷类化合物N在体外试验中对细胞株JEG-3、HepG2、细胞系HL-60的细胞毒活性； 2.JEG-3细胞、HL-60细胞体外经核苷类化合物N (0.1μM、0.2μM)处理24小时后,细胞经4%多聚甲醛固定及核染料Hoechst33258染色后,置荧光倒置显微镜下观察核苷类化合物N诱导细胞凋亡的形态学变化； 3.采用western blot技术,观察HL-60细胞经核苷类化合物N (0.1μM、0.2μM)处理后,核苷类化合物N对HL-60细胞凋亡有关蛋白caspase-3和AKT表达的调节作用。 结果 1.核苷类化合物N在体外能抑制HL-60细胞和JEG-3细胞的正常增殖,并表现出了明显的量效依赖关系； 2.不同浓度核苷类化合物N (0.1μM,0.2μM)处理HL-60细胞和JEG-3细胞24小时后,细胞经固定和Hoechst33258染色,荧光倒置显微镜观察发现,细胞细胞核形态发生明显改变,细胞核呈现染色不均,出现核切迹,核浓缩聚集,甚至细胞核碎裂。核苷类化合物N处理后细胞形态学表现出典型的凋亡改变； 3.体外核苷类化合物N (0.1μM、0.2μM)体外处理HL-60细胞24小时后,应用western blot技术,实验结果发现,核苷类化合物N可以明显下调HL-60细胞抗凋亡蛋白AKT的表达；经核苷类化合物N作用后,HL-60细胞出现caspase-3的裂解片段(17KD)。 结论 合成的核苷类化合物N在体外可以剂量依赖性地抑制多个肿瘤细胞株的生长繁殖,具有明显的细胞毒活性；核苷化合物N的抗肿瘤活性可能和其能调节肿瘤细胞凋亡相关蛋白表达,进而诱导肿瘤细胞凋亡有关。化合物N在体外试验中表现出了显著的抗肿瘤活性,该化合物的抗肿瘤活性和其具体抗肿瘤作用机制值得进一步深入开发研究。
[Abstract]:Objective to observe the antitumor activity of nucleoside compound N on several tumor cell lines in vitro by MTT and to explore the possible mechanism of the antitumor effect of the nucleoside compound. The results provide a theoretical basis for the development of new anti-tumor chemotherapeutic drugs or the discovery of lead compounds. Method 1. The cytotoxic activity of nucleoside compound N to HL-60 cell line JEG-3,HepG2, cell line was observed by conventional cytotoxicity test (MTT) in vitro. 2.JEG-3 cells and HL-60 cells were treated with N (0.1 渭 M) 0.2 渭 M for 24 hours in vitro. The cells were fixed with 4% paraformaldehyde and stained with nuclear dye Hoechst33258. The morphological changes of apoptosis induced by nucleoside compound N were observed under fluorescence inverted microscope. 3. Western blot technique was used to observe the regulatory effect of nucleoside N on the expression of caspase-3 and AKT in HL-60 cells treated with nucleoside compound N (0.1 渭 M 0.2 渭 M). Result 1. Nucleoside compound N could inhibit the normal proliferation of HL-60 cells and JEG-3 cells in vitro and showed a dose-dependent relationship. 2. After treated with different concentrations of nucleoside compound N (0.1 渭 M, 0.2 渭 M) for 24 hours, HL-60 cells and JEG-3 cells were fixed and stained with Hoechst33258, and observed by fluorescence inverted microscope. The nuclei showed uneven staining, nuclear cuts, nuclear condensation and aggregation, and even nuclear fragmentation. After treatment with nucleoside compound N, the cell morphology showed typical apoptotic changes. 3. Nuclear glycosides N (0. 1 渭 M + 0. 2 渭 M) were used to treat HL-60 cells for 24 hours. The results showed that nucleoside N could significantly down-regulate the expression of anti apoptotic protein AKT in HL-60 cells. After treatment with nucleoside compound N, caspase-3 fragmentation fragment (17KD) appeared in HL-60 cells. Conclusion the synthesized nucleoside compound N can inhibit the growth and reproduction of many tumor cell lines in a dose-dependent manner in vitro and has obvious cytotoxic activity. The antineoplastic activity of nucleoside N may be related to its ability to regulate the expression of apoptosis-related proteins in tumor cells and then induce apoptosis of tumor cells. Compound N showed significant antitumor activity in vitro, and its antitumor activity and its specific antitumor mechanism are worthy of further study.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96
本文编号：2390389
[Abstract]:Objective to observe the antitumor activity of nucleoside compound N on several tumor cell lines in vitro by MTT and to explore the possible mechanism of the antitumor effect of the nucleoside compound. The results provide a theoretical basis for the development of new anti-tumor chemotherapeutic drugs or the discovery of lead compounds. Method 1. The cytotoxic activity of nucleoside compound N to HL-60 cell line JEG-3,HepG2, cell line was observed by conventional cytotoxicity test (MTT) in vitro. 2.JEG-3 cells and HL-60 cells were treated with N (0.1 渭 M) 0.2 渭 M for 24 hours in vitro. The cells were fixed with 4% paraformaldehyde and stained with nuclear dye Hoechst33258. The morphological changes of apoptosis induced by nucleoside compound N were observed under fluorescence inverted microscope. 3. Western blot technique was used to observe the regulatory effect of nucleoside N on the expression of caspase-3 and AKT in HL-60 cells treated with nucleoside compound N (0.1 渭 M 0.2 渭 M). Result 1. Nucleoside compound N could inhibit the normal proliferation of HL-60 cells and JEG-3 cells in vitro and showed a dose-dependent relationship. 2. After treated with different concentrations of nucleoside compound N (0.1 渭 M, 0.2 渭 M) for 24 hours, HL-60 cells and JEG-3 cells were fixed and stained with Hoechst33258, and observed by fluorescence inverted microscope. The nuclei showed uneven staining, nuclear cuts, nuclear condensation and aggregation, and even nuclear fragmentation. After treatment with nucleoside compound N, the cell morphology showed typical apoptotic changes. 3. Nuclear glycosides N (0. 1 渭 M + 0. 2 渭 M) were used to treat HL-60 cells for 24 hours. The results showed that nucleoside N could significantly down-regulate the expression of anti apoptotic protein AKT in HL-60 cells. After treatment with nucleoside compound N, caspase-3 fragmentation fragment (17KD) appeared in HL-60 cells. Conclusion the synthesized nucleoside compound N can inhibit the growth and reproduction of many tumor cell lines in a dose-dependent manner in vitro and has obvious cytotoxic activity. The antineoplastic activity of nucleoside N may be related to its ability to regulate the expression of apoptosis-related proteins in tumor cells and then induce apoptosis of tumor cells. Compound N showed significant antitumor activity in vitro, and its antitumor activity and its specific antitumor mechanism are worthy of further study.
【学位授予单位】：新乡医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96
【参考文献】
相关期刊论文 前1条
1 余家华;李秋林;郑荣寿;张思维;葛莲英;曹骥;陈万青;;中国2009年喉恶性肿瘤发病和死亡分析[J];中国肿瘤;2013年05期
，本文编号：2390389
本文链接：https://www.wllwen.com/yixuelunwen/yiyaoxuelunwen/2390389.html
最近更新
教材专著
