14-噻吩基次亚甲基苦参碱体外抗鼻咽癌细胞CNE2作用的初步研究

发布时间：2018-05-04 14:48

本文选题：14-噻吩基次亚甲基苦参碱 + CNE2细胞　；参考：《广西医科大学》2011年硕士论文

【摘要】：背景和目的:鼻咽癌是南中国地区常见的头颈部肿瘤,治疗上推荐以放疗为主结合化疗的综合治疗方法,五年生存率可达50%以上。然而,不管是放射治疗或化学治疗均有一定的毒副作用,如何提高放化疗效果并减少放疗后并发症和减轻放疗的毒副作用是鼻咽癌研究关注的热点问题。由于肿瘤的远处转移和放射的局部损伤作用给进一步改善晚期鼻咽癌患者的预后和生活质量带来了困难,因此需要越来越多的化疗手段介入。苦参碱是近年受到关注的抗肿瘤成分,但存在生物利用率不高,并且有一定的毒副作用,使其难以单独用于临床抗肿瘤治疗。为了使苦参碱毒性降低和生物利用度更高,我们的合作者以苦参碱为先导化合物,通过化学合成方法对其结构进行改造,合成了20个结构新颖的苦参碱衍生物,并进行了抗鼻咽癌作用筛选。苦参碱改构体14-噻吩基次亚甲基苦参碱正是从中初步筛出的具有抗肿瘤活性的成分,本实验研究了该药对人鼻咽癌细胞CNE2体外增殖的影响,并探讨其机制。方法:选用鼻咽癌细胞株CNE2进行实验,首先经MTT进行抗鼻咽癌作用筛选;然后本实验分四组即是实验组、阴性对照组、正常细胞对照组、溶剂组、空白对照组(调零孔);倒置光学显微镜下观察细胞形态;MTT实验检测药物抑制率;流式细胞仪检测细胞周期的分布和凋亡情况。结果:加药细胞组形态变圆、变小、胞内可见空泡;MTT检测可见药物对人鼻咽癌细胞株CNE2有显著的抑制作用(P0.05),抑制率最大可达到97.9%,但是对人脐静脉内皮细胞没有表显出相应的抑制作用,抑制率最大仅达到7.7%;流式细胞仪检测结果可见CNE2细胞的周期阻滞在S期和有浓度依赖性的明显凋亡,凋亡率最大可达89.71%。结论:体外实验表明,14-噻吩基次亚甲基苦参碱具有明显的抗鼻咽癌作用,可通过细胞周期S期阻滞和诱导细胞凋亡来抑制鼻咽癌细胞的增殖分裂,并表现出很弱的毒性。
[Abstract]:Background and objective: nasopharyngeal carcinoma (NPC) is a common head and neck tumor in South China. Radiotherapy combined with chemotherapy is recommended. The 5-year survival rate is over 50%. However, both radiotherapy and chemotherapy have some side effects. How to improve the effect of radiotherapy and chemotherapy, reduce the complications after radiotherapy and reduce the side effects of radiotherapy is a hot issue in the research of nasopharyngeal carcinoma. Because the distant metastasis of the tumor and the local injury of radiation bring difficulties to further improve the prognosis and quality of life of the patients with advanced nasopharyngeal carcinoma, more and more chemotherapy interventions are needed. Matrine is an antitumor component that has attracted much attention in recent years, but its bioavailability is not high, and it has some toxic side effects, which makes it difficult to be used in clinical antitumor therapy alone. In order to reduce the toxicity and increase the bioavailability of matrine, our collaborators modified the structure of matrine by chemical synthesis method, and synthesized 20 novel matrine derivatives. The anti-nasopharyngeal carcinoma was screened. Matrine modified by 14 thienyl methylene matrine is a primary antitumor component. The effect of matrine on the proliferation of human nasopharyngeal carcinoma (NPC) cell line CNE2 in vitro was studied and its mechanism was discussed. Methods: the nasopharyngeal carcinoma cell line CNE2 was selected for the experiment, and the anti-nasopharyngeal carcinoma action was screened by MTT, then the experiment was divided into four groups: experimental group, negative control group, normal cell control group and solvent group. In the blank control group, the cell morphology and MTT assay were observed under inverted optical microscope, and the cell cycle distribution and apoptosis were detected by flow cytometry. Results: the morphology of the cells in the drug added group became round and small. MTT assay showed that the drug had a significant inhibitory effect on human nasopharyngeal carcinoma (NPC) cell line CNE2, with a maximum inhibition rate of 97.9%, but it had no apparent inhibitory effect on human umbilical vein endothelial cells (HUVEC). The inhibition rate was only 7.7.The results of flow cytometry showed that CNE2 cell cycle arrest was obvious in S phase and concentration-dependent manner, and the maximum apoptosis rate was 89.71%. Conclusion: the results of in vitro experiments showed that tetrathiophenyl methylene matrine could inhibit the proliferation and division of nasopharyngeal carcinoma cells through cell cycle S phase arrest and apoptosis, and showed a weak toxicity.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.63

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