心肌细胞培养液对鼻咽癌细胞cne-2增殖的影响及其机制的研究

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1、中图分类号：R89；R36；R39硕士学位论文心肌细胞培养液对鼻咽癌细胞CNE-2增殖的影响及其机制的研究院（系）、所临床医学院研究生姓名黄胜兰学科、专业肿瘤学导师姓名吴敬波教授二Ο一二年四月目录心肌细胞培养液对鼻咽癌细胞CNE-2增殖的影响及其机制的1研究„„„„„„„„„„„„„„„„„„„„„„„11.1中文摘要„„„„„„„„„„„„„„„„„„„„„21.2英文摘要„„„„„„„„„„„„„„„„„„„„„41.3前言„„„„„„„„„„„„„„„„„„„„„„„51.4材料与方法„„„„

2、„„„„„„„„„„„„„„„„121.5结果„„„„„„„„„„„„„„„„„„„„„„„211.6讨论„„„„„„„„„„„„„„„„„„„„„„„251.7结论„„„„„„„„„„„„„„„„„„„„„„„301.8参考文献„„„„„„„„„„„„„„„„„„„„„311.9英汉缩略词对照表„„„„„„„„„„„„„„„„„352致谢„„„„„„„„„„„„„„„„„„„„„„„363心肌细胞增殖的研究进展（综述）„„„„„„„„„„371心肌细胞培养液对鼻咽癌细胞CNE-2增殖的影响及其机制

3、的研究摘要本课题组前期研究体外已经证实心肌细胞培养液能明显抑制鼻咽癌细胞的生长，对非肿瘤细胞无抑制作用，推测心肌细胞培养液中可能含有一种或几种安全有效的抑瘤活性物质，该抑瘤活性无浓度依赖性。体内实验研究发现心肌细胞培养液能抑制S180小鼠移植瘤生长，不影响小鼠的正常生长；在CNE2裸鼠移植瘤模型得到了相同的答案，证实了该抑瘤活性物质的安全性和有效性，并进一步研究发现其抑瘤机制与诱导肿瘤细胞凋亡有关，同肿瘤微血管的生成无关；经葡聚糖凝胶层析分离提纯及质谱鉴定推测心肌细胞培养液中抑瘤活性成分中可能含有E

4、IF-5A或该蛋白的裂解片段。目的：通过体外抑瘤实验探讨心肌细胞培养液（myocardialcellsculturemedium，CMCM）对人鼻咽癌低分化鳞癌细胞系生长的影响，并对其抑瘤机制进行探讨。方法：分别于CMCM作用于CNE-2细胞12小时、24小时、36小时、48小时、60小时、72小时采用MTT法检测细胞的存活情况，同时以顺铂（DDP1.26μg/ml）作为阳性对照，RPMI1640培养基作为阴性对照，计算出细胞存活率，绘制生长曲线；并在作用24小时时采用流式细胞仪检测各组CNE-2细

5、胞的周期分布及凋亡情况。结果：作用12小时后CMCM组与DDP组细胞存活率（x±s，%）分别为87.27±6.99、89.88±0.72，与阴性对照组100.00±4.06相比便存在显著2差异（P<0.05）。随着时间的延长，差异逐渐增大，72小时后差异达最大，CMCM组与DDP组细胞存活率（x±s，%）分别为21.45±2.45、12.72±1.52，与阴性对照组100.00±10.69相比，差异均具有统计学意义。各实验组分别作用于鼻咽癌细胞24小时后，流式细胞仪检测细胞周期分布及凋亡情况。CMC

6、M组G0/G1期细胞比率（x±s，%）为68.70±3.40，与阴性对照组50.16±2.92相比，差异具有统计学意义，DDP组G0/G1期细胞比率为47.97±1.87，与阴性对照组相比，差异无统计学意义。DDP组G2/M期细胞比率（x±s，%）为22.10±2.67，明显高于阴性对照组6.60±2.03（P<0.05），CMCM组G2/M期细胞比率为8.10±1.78，与阴性对照组相比，差异无统计学意义。细胞凋亡率（x±s，%）CMCM组与DDP组分别为39.80±2.08、5.86±1.66，

7、与阴性对照组0.11±0.05相比，差异均具有统计学意义。结论：心肌细胞培养液对鼻咽癌细胞CNE-2生长的抑制作用具有时间依赖性。心肌细胞培养液鼻咽癌细胞CNE-2生长的抑制作用机制可能与诱导细胞凋亡及G0/G1期细胞阻滞有关。关键词：心肌细胞培养液；鼻咽癌细胞；时间依赖性；细胞周期；凋亡3Inhibitoryeffectsofmyocardialcellsculturemedium（CMCM)onnasopharyngealcarcinomacellsCNE-2andthepossiblemech

8、anismAbstract:bjective:Toinvestigatetheeffectofmyocardialcellsculturemedium（CMCM)ontheproliferationofhumannasopharyngealcarcinomacellsCNE-2andtorevealitsmechanism.Methods:ThesurvivalcellsofhumannasopharyngealcarcinomacellsCNE-2treatedwi