美洛昔康对人鼻咽癌CNE-1细胞株作用的研究
发布时间：2018-11-24 15:56
【摘要】： 目的初步探讨选择性环氧酶-2(COX-2)抑制剂美洛昔康(Meloxicam)对培养的人鼻咽癌CNE-1细胞增殖和凋亡的影响及其可能的机制,分析COX-2抑制剂在鼻咽癌化学预防和治疗中的应用价值,为今后的动物和临床研究提供可靠的实验依据。 方法选择CNE-1细胞为研究对象,以不同溶度(0、100μmol/L、200μmol/L、400μmol/L)美洛昔康处理CNE-1细胞不同时间(24h、48h、72h )后,用MTT法观察不同溶度、不同时间美洛昔康作用下CNE-1细胞的生长变化;将所选择药物作用于CNE-1细胞后,用AO+EB染色法、流式细胞仪检测法测定美洛昔康不同溶度作用下不同时间后细胞的凋亡情况;采用细胞免疫荧光法检测对照组和不同溶度美洛昔康作用48h后,CNE-1细胞内COX-2蛋白的表达情况。 结果 1.MTT比色法结果显示:美洛昔康对CNE-1细胞生长有显著的抑制作用,并且呈现出时间和剂量依赖性;随药物浓度加大和作用时间延长,存活细胞数量逐渐减少,在高溶度400umol/L时,48h后细胞生长抑制率高达50.3%;100μmol/L、200μmol/L、400μmol/L美洛昔康作用CNE-1细胞24h,其抑制率分别为8.4 %、10.3 %、16.4%;100μmol/L、200μmol/L、400μmol/L美洛昔康作用CNE-1细胞48h,其抑制率分别为10.4 %、28.8 %、50.3%; 2.AO+EB染色法及流式细胞检测法结果显示:用不同浓度的美洛昔康处理CNE-1细胞后,CNE-1细胞的凋亡率实验组与对照组相比均表现为不同程度的诱导作用(P0.05),并且这种诱导作用呈明显的剂量-时间依赖性。AO+EB染色结果可见明显的凋亡细胞存在,凋亡细胞数在高浓度美洛昔康组明显增多,400μmol/L溶度组干预48h后,200个细胞凋亡细胞达35±3个,流式细胞仪检查结果400μmol/L溶度组处理48h后细胞凋亡率为21.84±0.6%,与对照组有显著性差异(p0.05); 3.细胞免疫荧光结果显示:CNE-1细胞中COX-2蛋白表达呈阳性率高,美洛昔康作用后, CNE-1细胞中COX-2蛋白阳性的表达受到抑制,美洛昔康浓度越高,其抑制COX-2表达的作用越明显, 100μmol/L、200μmol/L、400μmol/L美洛昔康作用CNE-1细胞48h后,细胞内COX-2蛋白阳性表达率分别为39.61±0.34%、30.82±0.67%、23.03±1.12%,与对照组66.16±1.22%的阳性表达率有显著性差异(p0.05)。 结论 1.美洛昔康具有抑制CNE-1细胞增殖及诱导CNE-1细胞凋亡的作用,其抑制和诱导作用呈一定程度时间及剂量依赖性。 2.美洛昔康能抑制CNE-1细胞COX-2表达,其抑制作用呈时间依赖性。
[Abstract]:Objective to investigate the effects of meloxicam (Meloxicam), a selective cyclooxygenase-2 (COX-2) inhibitor, on proliferation and apoptosis of cultured CNE-1 cells from nasopharyngeal carcinoma (NPC) and its possible mechanism. To analyze the application value of COX-2 inhibitor in chemoprevention and treatment of nasopharyngeal carcinoma, and to provide reliable experimental basis for animal and clinical research in the future. Methods CNE-1 cells were treated with meloxicam (0100 渭 mol/L,200 渭 mol/L,400 渭 mol/L) for different time (24 h, 48 h, 72 h), and the different solubility was observed by MTT method. The growth and changes of CNE-1 cells under the action of meloxicam at different time; After the selected drugs were treated with CNE-1 cells, the apoptosis of CNE-1 cells was determined by AO EB staining and flow cytometry under different solubility of meloxicam. The expression of COX-2 protein in CNE-1 cells was detected by immunofluorescence assay after 48 hours of treatment with different solubility of meloxicam. Results the results of 1.MTT colorimetry showed that meloxicam significantly inhibited the growth of CNE-1 cells in a time-and dose-dependent manner. With the increase of drug concentration and the time of treatment, the number of viable cells gradually decreased. At the time of high solubility 400umol/L, the inhibition rate of cell growth was as high as 50.3% after 48 hours. The inhibitory rates of meloxicam on CNE-1 cells for 24 h were 8.4%, 10.3% and 16.4 渭 mol/L,200 渭 mol/L,400 渭 mol/L, respectively. The inhibitory rates of meloxicam on CNE-1 cells were 10.4%, 28.8% and 50.3% after 48 h treatment with 100 渭 mol/L,200 渭 mol/L,400 渭 mol/L. The results of 2.AO EB staining and flow cytometry showed that CNE-1 cells were treated with different concentrations of meloxicam. The apoptotic rate of CNE-1 cells in the experimental group was different from that in the control group (P0.05), and the apoptotic cells were found to exist in a dose-time dependent. AO EB staining. The number of apoptotic cells increased significantly in the high concentration meloxicam group. After 48 h intervention, the apoptosis rate of 200 cells reached 35 卤3 in the 400 渭 mol/L solubility group, and the apoptotic rate was 21.84 卤0.6 in the 400 渭 mol/L solubilization group after 48 h treatment by flow cytometry. There was significant difference between the control group and the control group (p0.05). 3. The results of cellular immunofluorescence showed that the positive rate of COX-2 protein in CNE-1 cells was high. After the treatment of meloxicam, the positive expression of COX-2 protein was inhibited. The higher the concentration of meloxicam was, the higher the expression of meloxicam was. The more obvious the inhibition of COX-2 expression was, the more obvious the positive expression rate of COX-2 protein in CNE-1 cells was 39.61 卤0.34and 30.82 卤0.673.03 卤1.12after treatment with meloxicam at 100 渭 mol/L,200 渭 mol/L,400 渭 mol/L for 48 h, respectively. There was significant difference between the positive rate of 66.16 卤1.22% and the control group (p0.05). Conclusion 1. Meloxicam can inhibit the proliferation of CNE-1 cells and induce apoptosis of CNE-1 cells in a time-and dose-dependent manner. 2. Meloxicam inhibited COX-2 expression in CNE-1 cells in a time dependent manner.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.63
本文编号：2354298
[Abstract]:Objective to investigate the effects of meloxicam (Meloxicam), a selective cyclooxygenase-2 (COX-2) inhibitor, on proliferation and apoptosis of cultured CNE-1 cells from nasopharyngeal carcinoma (NPC) and its possible mechanism. To analyze the application value of COX-2 inhibitor in chemoprevention and treatment of nasopharyngeal carcinoma, and to provide reliable experimental basis for animal and clinical research in the future. Methods CNE-1 cells were treated with meloxicam (0100 渭 mol/L,200 渭 mol/L,400 渭 mol/L) for different time (24 h, 48 h, 72 h), and the different solubility was observed by MTT method. The growth and changes of CNE-1 cells under the action of meloxicam at different time; After the selected drugs were treated with CNE-1 cells, the apoptosis of CNE-1 cells was determined by AO EB staining and flow cytometry under different solubility of meloxicam. The expression of COX-2 protein in CNE-1 cells was detected by immunofluorescence assay after 48 hours of treatment with different solubility of meloxicam. Results the results of 1.MTT colorimetry showed that meloxicam significantly inhibited the growth of CNE-1 cells in a time-and dose-dependent manner. With the increase of drug concentration and the time of treatment, the number of viable cells gradually decreased. At the time of high solubility 400umol/L, the inhibition rate of cell growth was as high as 50.3% after 48 hours. The inhibitory rates of meloxicam on CNE-1 cells for 24 h were 8.4%, 10.3% and 16.4 渭 mol/L,200 渭 mol/L,400 渭 mol/L, respectively. The inhibitory rates of meloxicam on CNE-1 cells were 10.4%, 28.8% and 50.3% after 48 h treatment with 100 渭 mol/L,200 渭 mol/L,400 渭 mol/L. The results of 2.AO EB staining and flow cytometry showed that CNE-1 cells were treated with different concentrations of meloxicam. The apoptotic rate of CNE-1 cells in the experimental group was different from that in the control group (P0.05), and the apoptotic cells were found to exist in a dose-time dependent. AO EB staining. The number of apoptotic cells increased significantly in the high concentration meloxicam group. After 48 h intervention, the apoptosis rate of 200 cells reached 35 卤3 in the 400 渭 mol/L solubility group, and the apoptotic rate was 21.84 卤0.6 in the 400 渭 mol/L solubilization group after 48 h treatment by flow cytometry. There was significant difference between the control group and the control group (p0.05). 3. The results of cellular immunofluorescence showed that the positive rate of COX-2 protein in CNE-1 cells was high. After the treatment of meloxicam, the positive expression of COX-2 protein was inhibited. The higher the concentration of meloxicam was, the higher the expression of meloxicam was. The more obvious the inhibition of COX-2 expression was, the more obvious the positive expression rate of COX-2 protein in CNE-1 cells was 39.61 卤0.34and 30.82 卤0.673.03 卤1.12after treatment with meloxicam at 100 渭 mol/L,200 渭 mol/L,400 渭 mol/L for 48 h, respectively. There was significant difference between the positive rate of 66.16 卤1.22% and the control group (p0.05). Conclusion 1. Meloxicam can inhibit the proliferation of CNE-1 cells and induce apoptosis of CNE-1 cells in a time-and dose-dependent manner. 2. Meloxicam inhibited COX-2 expression in CNE-1 cells in a time dependent manner.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R739.63
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