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目的研究三氧化二砷(As2O3)诱导人鼻咽癌细胞株凋亡作用及其相关机制。方法采用流式细胞术、电镜、TUNEL方法检测As2O3诱导CNE1细胞凋亡作用,免疫组织化学法检测As2O3对CNE1细胞p53、bcl-2和bax蛋白表达的影响。结果经As2O3处理的CNE1细胞发生凋亡,表现为FCM可检测到“亚二倍体峰”,形态学上可见核固缩,染色质边集,凋亡小体形成等改变;TUNEL法可检测到细胞内呈棕色颗粒的凋亡细胞,随着药物浓度升高,凋亡细胞发生率逐渐增多,As2O3目浓度为0.5mg/L、1.0mg/L和2.0mg/L作用48h后,CNE1细胞的凋亡指数分别为2.66±0.64、8.15±0.96和11.59±0.68,显著高于非药物处理组(0.43±0.43,P<0.05)。经As2O3处理的CNE1细胞内p53和bax蛋白表达较对照组明显增加,与凋亡指数呈正相关关系(r=0.554,P=0.011;r=0.891,P=0.000…);p53和bax蛋白表达也呈正相关关系(r=0.626,P=0.003)。结论As2O3在体外可诱导鼻咽癌CNE1细胞凋亡,其机制可能与上调p53、bax基因表达有关。
Objective To study the apoptosis of human nasopharyngeal carcinoma cell line induced by As2O3 and its related mechanism. Methods Flow cytometry, electron microscopy and TUNEL were used to detect the apoptosis of CNE1 cells induced by As2O3. The effects of As2O3 on the expression of p53, bcl-2 and bax proteins were detected by immunohistochemistry. Results Apoptosis of CNE1 cells treated with As2O3 showed that the subduploid peak was detected by FCM. Nuclear condensation, chromatin margination and formation of apoptotic bodies were observed by TUNEL assay. The number of apoptotic cells increased gradually with the increase of drug concentration. After the concentration of As2O3 was 0.5mg / L, 1.0mg / L and 2.0mg / L for 48h, the number of CNE1 cells The apoptosis index was 2.66 ± 0.64, 8.15 ± 0.96 and 11.59 ± 0.68 respectively, which was significantly higher than that of non-drug treatment group (0.43 ± 0.43, P <0.05). The expression of p53 and bax in As2O3-treated CNE1 cells was significantly increased compared with the control group, and positively correlated with the apoptosis index (r = 0.554, P = 0.011; r = 0.891, P = 0.000, There was a positive correlation (r = 0.626, P = 0.003). Conclusion As2O3 can induce CNE1 cell apoptosis in vitro, and its mechanism may be related to the up-regulation of p53 and bax gene expression.