海带多糖对人鼻咽癌细胞及其裸鼠移植瘤抑制作用的初步研究

发布时间：2018-05-04 20:43

本文选题：鼻咽癌细胞系 + 海带多糖　；参考：《广西医科大学》2012年硕士论文

【摘要】：目的鼻咽癌(nasopharyngeal carcinoma,NPC)是中国南方地区最常见的头颈部恶性肿瘤,主要治疗方法是以放射治疗为主辅以化学药物治疗的综合治疗,而化学药物治疗对中晚期的鼻咽癌患者尤为重要,但治疗剂量的化疗药物多有较多严重的毒副作用。海带多糖(Laminaria Japonica Polysaccharides,LJP)是从海带中分离提取出的具有生物活性的植物多糖,具有抗肿瘤、提高免疫功能、防辐射等多种活性功能,本研究旨在通过观察海带多糖在体内外对人鼻咽癌细胞的抑制作用,探讨其可能的抗癌机制。方法运用MTT法检测不同浓度的海带多糖对人鼻咽癌细胞株(HONE1和CNE2)增殖的抑制作用。运用Annexin V-FITC/PI双染法检测海带多糖对HONE1细胞凋亡的影响。以人鼻咽癌细胞株HONE1建立裸鼠皮下移植瘤模型,将造模成功的裸鼠随机分入5个处理组：生理盐水组、顺铂组、海带多糖低、中、高剂量组,每组6只,观察各组移植瘤体积和裸鼠体重的变化情况。2周治疗结束后,处死裸鼠,剖出瘤块,称瘤重并计算各组的抑瘤率,并在透射电镜(Transmission electron microscopy,TEM)观察移植瘤细胞超微结构的改变。结果MTT结果显示,海带多糖对鼻咽癌细胞(HONE1和CNE2)的增殖有抑制作用,且具有剂量相关性,320mg/L的海带多糖作用72h的抑制率分别为58.27%(P0.01)和55.00%(P0.01)；流式细胞术(Flow Cytometry, FCM)检测结果表明,海带多糖具有诱导HONE1细胞凋亡的作用,凋亡率随着海带多糖浓度的增加而增高,320mg/L的凋亡率为(49.51±1.89)%(P0.01)。体内实验中,海带多糖中、高剂量组抑瘤率分别为33.7%(P0.05)和47.0%(P0.01),具有明显抑制移植瘤的作用,而低剂量组的抑瘤率仅为16.4%(P0.05)。透射电镜结果提示移植瘤组织的癌细胞呈现凋亡独特的改变：细胞皱缩、染色质凝聚、胞核碎裂、胞浆空泡变性、胞膜起泡、形成游离的凋亡小体。结论海带多糖具有抑制鼻咽癌细胞生长的作用,机制可能是通过诱导癌细胞凋亡而实现的。
[Abstract]:Objective nasopharyngeal carcinoma (NPC) is the most common malignant tumor of head and neck in southern China. The main treatment method is radiotherapy combined with chemotherapeutic therapy, and chemotherapy is especially important for patients with advanced nasopharyngeal carcinoma. However, the therapeutic dose of chemotherapeutic drugs have more serious side effects. Laminaria Japonica Polysaccharide (Laminaria Japonica Polysaccharide) is a bioactive plant polysaccharide isolated from Laminaria japonica. The aim of this study was to observe the inhibitory effect of kelp polysaccharides on human nasopharyngeal carcinoma cells in vitro and in vivo, and to explore its possible anticancer mechanism. Methods the inhibitory effects of different concentrations of kelp polysaccharides on the proliferation of human nasopharyngeal carcinoma cell lines (HONE1 and CNE2) were detected by MTT assay. The effect of kelp polysaccharide on apoptosis of HONE1 cells was detected by Annexin V-FITC/PI double staining. Human nasopharyngeal carcinoma (NPC) cell line HONE1 was used to establish subcutaneous transplanted tumor model in nude mice. The successful nude mice were randomly divided into 5 groups: normal saline group, cisplatin group, kelp polysaccharide low, middle and high dose group, each group with 6 rats. After 2 weeks of treatment, nude mice were killed, tumor weight was weighed, tumor inhibition rate was calculated, and ultrastructural changes of transplanted tumor cells were observed under transmission electron microscope (TEM). Results MTT results showed that kelp polysaccharides could inhibit the proliferation of nasopharyngeal carcinoma cells (HONE1 and CNE2), and the inhibitory rates of kelp polysaccharides with dose-dependent manner were 58.27mg / L and 55.00% P0.01respectively, and the flow Cytometry (FCM) showed that the inhibitory rates of kelp polysaccharides were 58.27mg / L and 55.00g / L, respectively, and the results of flow Cytometry (FCM) showed that Kelp polysaccharides could induce apoptosis of HONE1 cells. The apoptotic rate increased with the increase of kelp polysaccharide concentration, and the apoptotic rate of 320mg / L was 49.51 卤1.89g / L. In vivo experiments, the inhibitory rates of high dose kelp polysaccharides were 33.7 and 47.0, respectively, which could inhibit the transplanted tumor obviously, but in the low dose group, the tumor inhibition rate was only 16.4% and P0.05%. The results of transmission electron microscope showed that the cancer cells of transplanted tumor showed unique changes of apoptosis, such as cell shrinkage, chromatin condensation, nuclear fragmentation, cytosolic vacuolar degeneration, membrane bubbling, and free apoptotic bodies. Conclusion Laminaria polysaccharide can inhibit the growth of nasopharyngeal carcinoma cells, and the mechanism may be by inducing apoptosis of cancer cells.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.63

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