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目的应用白藜芦醇作用于咽鳞癌细胞株FADU,观察其放疗增敏作用。方法体外培养咽鳞癌细胞株FADU,以细胞毒实验(cytotoxicity test,MTT)检测不同浓度白藜芦醇对细胞增殖抑制情况;用克隆形成法绘制细胞存活曲线,获得白藜芦醇的放射增敏比(sensitive enhancement ratio,SER)。以流式细胞术(flow cytometry,FCM)分析对照组、单纯照射组、白藜芦醇组和照射增敏组(白藜芦醇联合照射组)细胞周期变化和凋亡情况。结果 MTT结果显示白藜芦醇对FADU细胞生长抑制作用随药物浓度的增加而增强(P<0.05)。克隆形成实验提示单纯照射组在2Gy时细胞存活分数(surviving fraction at 2Gy,SF2)为0.717±0.062;照射增敏组SF2为0.426±0.035,SER为1.684±0.178,差异有统计学意义(P=0.007)。FCM显示FADU细胞经4Gy照射后,G_2/M期细胞比例增加,G_1期细胞比例减少;经白藜芦醇作用24 h后,G_1期细胞比例减少,G_2/M期和S期细胞比例增加;当白藜芦醇与4 Gy照射联合应用时,G_2/M期细胞比例显著增加,G_1期细胞比例显著减少。对照组、单纯照射组、白藜芦醇组和照射增敏组FADU细胞的凋亡率(%)分别为1.94±1.65、4.56±0.92、2.03±1.46及23.1 1±7.22,照射增敏组凋亡率与其他各处理组比较,差异均有统计学意义(P<0.05)。结论白藜芦醇可增强咽鳞癌细胞株FADU放射敏感性,其机制可能与改变细胞周期的分布、诱导细胞凋亡相关。
Objective To investigate the radiosensitization effect of resveratrol on pharyngeal squamous cell carcinoma cell line FADU. Methods The pharyngeal squamous cell carcinoma cell line FADU was cultured in vitro. The inhibitory effect of different concentrations of resveratrol on the cell proliferation was detected by cytotoxicity test (MTT). The cell viability curve was drawn by clone formation method to obtain the radioactive increase of resveratrol Sensitive enhancement ratio (SER). The changes of cell cycle and apoptosis in control group, radiation group, resveratrol group and irradiation sensitization group (resveratrol combined irradiation group) were analyzed by flow cytometry (FCM). Results MTT results showed that resveratrol inhibited the growth of FADU cells with the increase of drug concentration (P <0.05). The clonogenic assay indicated that the surviving fraction at 2Gy was 0.717 ± 0.062 at 2 Gy in SFI group, SF2 was 0.426 ± 0.035 and SER was 1.684 ± 0.178 in irradiation-sensitized group, the difference was statistically significant (P = 0.007). FCM showed that the proportion of cells in G_2 / M phase was increased and the proportion of cells in G_1 phase decreased after irradiated with 4Gy for FADU cells. The proportion of cells in G_1 phase decreased and the proportion of cells in G_2 / M phase and S phase increased after resveratrol treatment for 24 h. When resveratrol was used in combination with 4 Gy, the proportion of cells in G 2 / M phase increased significantly, and the proportion of cells in G 1 phase decreased significantly. The apoptotic rates (%) of FADU cells in control, irradiation group, resveratrol group and irradiation sensitized group were 1.94 ± 1.65, 4.56 ± 0.92, 2.03 ± 1.46 and 23.1 1 ± 7.22, respectively Mortality and other treatment groups, the differences were statistically significant (P <0.05). Conclusion Resveratrol can enhance radiosensitivity of pharyngeal squamous cell carcinoma cell line FADU, and its mechanism may be related to changing the distribution of cell cycle and inducing cell apoptosis.