血管内皮抑制素联合放疗对人鼻咽癌细胞CNE-2的体外实验研究

发布时间：2018-06-23 19:19

本文选题：重组人血管内皮抑制素(恩度) + 放疗增敏　；参考：《昆明医学院》2010年硕士论文

【摘要】： 【目的】观察血管内皮抑制素(恩度)联合放疗对人鼻咽癌细胞系(CNE2)的增殖抑制和诱导细胞凋亡的作用,及其对细胞周期的影响。为血管内皮抑制素对鼻咽癌放疗增敏的可行性提供实验依据。【方法】四噻唑蓝比色试验(MTT)观察不同浓度的血管内皮抑制素对鼻咽癌CNE2浓度时间抑制作用,计算出最适浓度。四噻唑蓝比色试验(MTT)法观察血管内皮抑制素联合放疗对鼻咽癌CNE2时间抑制作用,以流式细胞术检测对其细胞凋亡和周期的影响。【结果】1.血管内皮抑制素处理后,用倒置显微镜观察恩度处理后的鼻咽癌细胞,发现随药物及作用时间的增加,CNE2癌细胞逐渐变圆浮起,折光度降低,细胞间连接松散,增殖减慢。 2.MTT法检测不同浓度的恩度在不同时间点对CNE2的增殖抑制作用。不同浓度恩度作用于鼻咽癌细胞CNE2,随着抑制剂作用浓度增加、作用时间的延长,肿瘤细胞存活率逐步降低。 3.单次放疗剂量4Gy,恩度在浓度100mg/1的时候,综合治疗对鼻咽癌细胞CNE2的综合作用强,综合作用比单独用药或者放疗作用强,随时间的增加,肿瘤细胞的存活率逐步降低。 4.流式细胞分析显示：人鼻咽癌细胞CNE2胞凋亡率与G2/M期百分率呈负相关,恩度可能增加了细胞G2/M的放射敏感性。 5.TdT介导的dUTP缺口末端标记技术(TUNEL)法检测细胞凋亡：恩度浓度为100mg/1,单次剂量4Gy,作用72h,以PI双染色法流式细胞仪测定细胞凋亡率和坏死率,对照组细胞凋亡率仅为1.7%,细胞坏死率为8.1%,实验组细胞凋亡率为11.6%。【结论】1.一定浓度的血管内皮抑制素对人鼻咽癌细胞系(CNE2)有抑制作用,作用呈时间浓度依耐性。 2.血管内皮抑制素联合放疗对人鼻咽癌细胞系(CNE2)有增敏作用,呈时间依耐性。
[Abstract]:[objective] to observe the effects of vascular endothelial inhibitor (Endothelin) combined with radiotherapy on the proliferation and apoptosis of human nasopharyngeal carcinoma cell line (CNE2) and its effect on cell cycle. [methods] MTT assay was used to observe the time inhibitory effect of different concentrations of vascular endothelin on CNE2 concentration in nasopharyngeal carcinoma (NPC). The optimum concentration is calculated. MTT assay was used to observe the inhibitory effect of vascular endothelial inhibitor combined with radiotherapy on CNE2 of nasopharyngeal carcinoma, and the effect of flow cytometry on apoptosis and cell cycle of nasopharyngeal carcinoma. [results] 1. After treatment with vascular endothelin, nasopharyngeal carcinoma cells treated with Endosterone were observed by inverted microscope. With the increase of drug and time of action, CNE2 cancer cells gradually became round and floating, the diopter decreased, and the intercellular junctions were loosely connected. 2. MTT assay was used to detect the inhibitory effect of different concentrations of Endox on CNE2 at different time points. The survival rate of nasopharyngeal carcinoma cell line CNE _ 2 decreased gradually with the increase of the concentration of inhibitor and the prolongation of the action time. When the dose of single dose of radiotherapy was 4 Gy, the comprehensive effect of combined therapy on nasopharyngeal carcinoma cell line CNE2 was stronger than that of single dose of radiotherapy or radiotherapy at the concentration of 100mg/1, and the effect increased with time. The survival rate of tumor cells decreased gradually. 4. 4. Flow cytometry showed that the apoptosis rate of CNE2 cells was negatively correlated with the percentage of G _ 2 / M phase. TDT mediated dUTP Nick end labeling (Tunel) assay was used to detect apoptosis: Endol concentration was 100 mg / 1, single dose was 4 Gy, contact time was 72 h, flow fine flow was detected by Pi double staining. Cell apoptosis rate and necrosis rate were measured by cell analyzer. In the control group, the apoptosis rate was only 1.7%, the necrosis rate was 8.1%, and the apoptotic rate in the experimental group was 11.6%. [conclusion] 1. A certain concentration of vascular endothelin inhibited human nasopharyngeal carcinoma (CNE2) cell line (CNE2), the effect showed time concentration dependent tolerance. Endotheliostatin combined with radiotherapy can enhance the sensitivity of human nasopharyngeal carcinoma cell line (CNE2) with time dependent tolerance.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.63

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