【摘要】：目的 本實驗旨在探討去甲斑蝥酸鈉(Disodium Cantharidinate)對鼻咽癌CNE-2細(xì)胞株體外的放射增敏作用,并初步探討其機(jī)理。 方法 應(yīng)用MTS法測定不同時間、不同濃度去甲斑蝥素鈉對鼻咽癌CNE-2細(xì)胞的增殖抑制作用,求出半數(shù)抑制濃度(50%Inhibiting Concentration, IC50)。將鼻咽癌CNE-2細(xì)胞分為：空白對照組、照射組、藥物組、藥物與照射聯(lián)合組,應(yīng)用克隆形成法研究去甲斑蝥酸鈉與放射聯(lián)合效應(yīng),利用“多靶單擊”數(shù)學(xué)模型[SF=1-(1-e-D/D0)ΔN進(jìn)行曲線擬合,獲得細(xì)胞存活曲線及平均致死劑量、準(zhǔn)閾劑量、放射增敏比(Sensitization enhancement ratio, SER)等參數(shù),進(jìn)行效應(yīng)評估。將鼻咽癌CNE-2細(xì)胞分為：空白對照組、照射組、藥物組、藥物與照射聯(lián)合組,去甲斑蝥酸鈉作用細(xì)胞48h后,流式細(xì)胞術(shù)分析細(xì)胞凋亡率及周期的變化。 結(jié)果 去甲斑蝥酸鈉作用后鼻咽癌CNE-2細(xì)胞生長增殖受到抑制,作用48小時IC50為340mg/L。20mg/L去甲斑蝥酸鈉作用鼻咽癌CNE-2細(xì)胞48h后聯(lián)合放射的放射增敏比(SER)為1.43。去甲斑蝥酸鈉與放療作用于CNE-2細(xì)胞株后流式細(xì)胞儀檢測細(xì)胞周期,G2/M期細(xì)胞急劇增高,由8.26%上升至48.3%,G1和S期細(xì)胞明顯減少。CNE-2細(xì)胞經(jīng)4Gy照射、20mg/L去甲斑蝥酸鈉處理后,凋亡率較空白對照組細(xì)胞升高,分別為12.68%,5.37%,而照射聯(lián)合去甲斑蝥酸鈉處理組中,凋亡率升高更為顯著,可達(dá)18.84%。比較差異均有統(tǒng)計學(xué)意義(P0.05)。 結(jié)論 1、去甲斑蝥酸鈉對體外培養(yǎng)的鼻咽癌CNE-2細(xì)胞株有增殖抑制作用,其作用與藥物濃度、作用時間呈正相關(guān)性。 2、去甲斑蝥酸鈉對體外培養(yǎng)的鼻咽癌CNE-2細(xì)胞株有放射增敏作用。 3、去甲斑蝥酸鈉增加鼻咽癌CNE-2細(xì)胞放射敏感性的可能機(jī)制是阻滯細(xì)胞周期、誘導(dǎo)細(xì)胞凋亡。
[Abstract]:Objective to investigate the radiosensitizing effect of sodium norcantharidate (Disodium Cantharidinate) on nasopharyngeal carcinoma (NPC) CNE-2 cell line in vitro and its mechanism. Methods the inhibitory effects of norcantharidin sodium at different concentrations on the proliferation of nasopharyngeal carcinoma (NPC) CNE-2 cells were determined by MTS assay. The half inhibitory concentration (50%Inhibiting Concentration, IC50) was calculated. Nasopharyngeal carcinoma (NPC) CNE-2 cells were divided into three groups: blank control group, irradiation group, combined drug and irradiation group. The combined effect of sodium norcantharidate and radiation was studied by clone formation method. The curve was fitted by SF=1- (1-e-D/D0) 螖 N, a mathematical model of "multi-target click". Cell survival curve, average lethal dose, quasi-threshold dose and radiosensitization ratio (Sensitization enhancement ratio, SER) were obtained to evaluate the effects. Nasopharyngeal carcinoma (NPC) CNE-2 cells were divided into three groups: blank control group, irradiation group, drug and irradiation combined group. After 48 hours of treatment with sodium norcantharidate, apoptosis rate and cell cycle were analyzed by flow cytometry. Results the growth and proliferation of nasopharyngeal carcinoma CNE-2 cells were inhibited after treated with sodium norcantharidate. The radiosensitization ratio (SER) of 340mg/L.20mg/L norcantharidate sodium to 340mg/L.20mg/L norcantharidate sodium for 48 h was 1.43. After treated with sodium norcantharidate and radiotherapy in CNE-2 cell line, the cell cycle was detected by flow cytometry. The cell cycle of G 2 / M phase increased sharply from 8.26% to 48.3%. CNE-2 cells were irradiated by 4Gy and treated with 20mg/L sodium norcantharidate. The apoptotic rate was higher than that in the blank control group (12.68 and 5.37, respectively), while the apoptosis rate in the irradiation combined with sodium norcantharidate group was higher than that in the blank control group (18.84%). The difference was statistically significant (P0.05). Conclusion 1. Sodium norcantharidate can inhibit the proliferation of nasopharyngeal carcinoma (NPC) CNE-2 cell line in vitro. 2. Sodium norcantharidate has radiosensitizing effect on CNE-2 cell line of nasopharyngeal carcinoma in vitro. 3. The possible mechanism of sodium norcantharidate to increase radiosensitivity of CNE-2 cell line of nasopharyngeal carcinoma To block the cell cycle, Induce apoptosis.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R739.63

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