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

发布时间：2018-06-19 08:52

本文选题：心肌细胞培养液 + 鼻咽癌细胞　；参考：《泸州医学院》2012年硕士论文

【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EIF-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（myocardial cells culturemedium，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相比便存在显著差异（P0.05）。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。CMCM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（P0.05），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，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。
[Abstract]:Our previous studies have demonstrated in vitro that cardiomyocyte culture medium can significantly inhibit the growth of nasopharyngeal carcinoma cells, but has no inhibitory effect on non-tumor cells. It is speculated that there may be one or several safe and effective anti-tumor active substances in cardiomyocyte culture medium, which is not concentration-dependent. In vivo experiments showed that cardiomyocyte culture medium could inhibit the growth of transplanted tumor in S180 mice without affecting the normal growth of mice. The same answer was obtained in CNE2 nude mice model of transplanted tumor, which confirmed the safety and efficacy of the anti-tumor active substance. Furthermore, it was found that the mechanism of tumor inhibition was related to the induction of apoptosis of tumor cells, but not to the formation of tumor microvessels. The anti-tumor components of cardiomyocyte culture medium may contain the fragment of EIF-5A or this protein in the cardiomyocyte culture medium by Glucan gel chromatography separation and mass spectrometry. Objective: to investigate the effect of myocardial cells culturemedia (CMCM) on the growth of poorly differentiated human nasopharyngeal carcinoma (NPC) cell lines in vitro and its mechanism. Methods: CNE-2 cells were treated with CMCM for 12 hours, 24 hours, 36 hours, 48 hours, 60 hours and 72 hours, respectively. MTT assay was used to detect the survival rate of CNE-2 cells, and cisplatin DDP 1.26 渭 g / ml was used as negative control to calculate the survival rate of CNE-2 cells. The growth curve was drawn and the cell cycle distribution and apoptosis of CNE-2 cells in each group were detected by flow cytometry at 24 hours. Results: the cell survival rate of CMCM group and DDP group was 87.27 卤6.9 卤89.88 卤0.72, which was significantly higher than that of negative control group (100.00 卤4.06). With the prolongation of time, the difference gradually increased 72 hours later, the difference reached the maximum after 72 hours. The cell survival rate of CMCM group and DDP group were 21.45 卤2.45 卤1.52 卤1.52 and 100.00 卤10.69, respectively. The difference was statistically significant compared with that of negative control group (100.00 卤10.69). After treated with nasopharyngeal carcinoma cells for 24 hours, the cell cycle distribution and apoptosis were detected by flow cytometry. In CMCM group, the cell ratio of G _ 0 / G _ 1 phase was 68.70 卤3.40, which was higher than that of negative control group (50.16 卤2.92). The ratio of G _ 2 / M phase cells in the DDP group was 22.10 卤2.67, which was significantly higher than that in the negative control group (6.60 卤2.03P0.05CMCM, 8.10 卤1.78), compared with the negative control group, the ratio of G _ 2 / M phase cells in the DDP group was significantly higher than that in the negative control group (47.97 卤1.87), and the ratio of G _ 2 / M phase cells in the DDP group was 22.10 卤2.67, which was significantly higher than that in the negative control group (8.10 卤1.78). The difference was not statistically significant. The rate of apoptosis in CMCM group and DDP group was 39.80 卤2.080.86 卤1.66, respectively, which was significantly higher than that in negative control group (0.11 卤0.05). Conclusion: the inhibitory effect of cardiomyocyte culture medium on the growth of nasopharyngeal carcinoma cell line CNE-2 is time dependent. The inhibitory mechanism of CNE-2 growth in cardiomyocyte culture medium may be related to the induction of apoptosis and the arrest of G _ 0 / G _ 1 phase cells.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.63

【参考文献】