美洛昔康對人鼻咽癌CNE-1細胞株作用的研究
發(fā)布時間：2018-11-24 15:56
【摘要】： 目的初步探討選擇性環(huán)氧酶-2(COX-2)抑制劑美洛昔康(Meloxicam)對培養(yǎng)的人鼻咽癌CNE-1細胞增殖和凋亡的影響及其可能的機制,分析COX-2抑制劑在鼻咽癌化學預防和治療中的應(yīng)用價值,為今后的動物和臨床研究提供可靠的實驗依據(jù)。 方法選擇CNE-1細胞為研究對象,以不同溶度(0、100μmol/L、200μmol/L、400μmol/L)美洛昔康處理CNE-1細胞不同時間(24h、48h、72h )后,用MTT法觀察不同溶度、不同時間美洛昔康作用下CNE-1細胞的生長變化;將所選擇藥物作用于CNE-1細胞后,用AO+EB染色法、流式細胞儀檢測法測定美洛昔康不同溶度作用下不同時間后細胞的凋亡情況;采用細胞免疫熒光法檢測對照組和不同溶度美洛昔康作用48h后,CNE-1細胞內(nèi)COX-2蛋白的表達情況。 結(jié)果 1.MTT比色法結(jié)果顯示:美洛昔康對CNE-1細胞生長有顯著的抑制作用,并且呈現(xiàn)出時間和劑量依賴性;隨藥物濃度加大和作用時間延長,存活細胞數(shù)量逐漸減少,在高溶度400umol/L時,48h后細胞生長抑制率高達50.3%;100μmol/L、200μmol/L、400μmol/L美洛昔康作用CNE-1細胞24h,其抑制率分別為8.4 %、10.3 %、16.4%;100μmol/L、200μmol/L、400μmol/L美洛昔康作用CNE-1細胞48h,其抑制率分別為10.4 %、28.8 %、50.3%; 2.AO+EB染色法及流式細胞檢測法結(jié)果顯示:用不同濃度的美洛昔康處理CNE-1細胞后,CNE-1細胞的凋亡率實驗組與對照組相比均表現(xiàn)為不同程度的誘導作用(P0.05),并且這種誘導作用呈明顯的劑量-時間依賴性。AO+EB染色結(jié)果可見明顯的凋亡細胞存在,凋亡細胞數(shù)在高濃度美洛昔康組明顯增多,400μmol/L溶度組干預48h后,200個細胞凋亡細胞達35±3個,流式細胞儀檢查結(jié)果400μmol/L溶度組處理48h后細胞凋亡率為21.84±0.6%,與對照組有顯著性差異(p0.05); 3.細胞免疫熒光結(jié)果顯示:CNE-1細胞中COX-2蛋白表達呈陽性率高,美洛昔康作用后, CNE-1細胞中COX-2蛋白陽性的表達受到抑制,美洛昔康濃度越高,其抑制COX-2表達的作用越明顯, 100μmol/L、200μmol/L、400μmol/L美洛昔康作用CNE-1細胞48h后,細胞內(nèi)COX-2蛋白陽性表達率分別為39.61±0.34%、30.82±0.67%、23.03±1.12%,與對照組66.16±1.22%的陽性表達率有顯著性差異(p0.05)。 結(jié)論 1.美洛昔康具有抑制CNE-1細胞增殖及誘導CNE-1細胞凋亡的作用,其抑制和誘導作用呈一定程度時間及劑量依賴性。 2.美洛昔康能抑制CNE-1細胞COX-2表達,其抑制作用呈時間依賴性。
[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 鹵3 in the 400 渭 mol/L solubility group, and the apoptotic rate was 21.84 鹵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 鹵0.34and 30.82 鹵0.673.03 鹵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 鹵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.
【學位授予單位】：南華大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R739.63
本文編號：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 鹵3 in the 400 渭 mol/L solubility group, and the apoptotic rate was 21.84 鹵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 鹵0.34and 30.82 鹵0.673.03 鹵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 鹵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.
【學位授予單位】：南華大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R739.63
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