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背景与目的:有关鼻咽癌的生物节律特征尚未见文献报道。本文拟探索荷人鼻咽癌裸鼠骨髓细胞的DNA合成及几种凋亡相关基因表达的生物节律,为临床制定鼻咽癌时辰化疗方案提供必要的参数。方法:BALB/C裸小鼠69只,置于程控的独立光照系统(12h光照,12h黑暗)同步化至少3周。然后双侧腋窝皮下接种人鼻咽低分化鳞癌细胞(CNE-2)裸小鼠移植瘤,成瘤后按6个不同时间点分批处死小鼠,收集骨髓细胞,固定、染色后流式细胞仪测DNA含量,用ANOVA法检验各期细胞在6个时间点差异的显著性,Cosinor法考察G1、S、G2/M期细胞在24h的分布是否符合余弦函数。部分骨髓细胞经裂解、蛋白定量后用WesternBlot法检测不同时间点p53、p21和Bcl-2三种基因蛋白表达的变化情况。结果:G1、S、G2/M期细胞在3、7、11、15、19、23HALO(HoursAfterLightOnset即灯亮后第3、7、11、19、23h)的分布随时间变化有显著性差异,G1、G2/M期细胞24h变化符合余弦节律,高峰值分别位于10.8HALO和1.8HALO。骨髓细胞p53和Bcl-2蛋白表达24h强度不同。p53在11HALO最强,在7HALO最弱;Bcl-2在15HALO最强,在19HALO最弱,p21蛋白未见表达。结论:荷人鼻咽癌裸小鼠骨髓细胞DNA合成随昼夜交替呈节律变化,p53和Bcl-2蛋白表达随昼夜时间变化而规律性波动。
BACKGROUND & OBJECTIVE: The biological rhythms of nasopharyngeal carcinoma have not been reported in the literature. This article intends to explore the DNA synthesis of human nasopharyngeal carcinoma nude mouse bone marrow cells and several apoptosis-related gene expression of the biological rhythm for the development of clinical chemotherapy for nasopharyngeal carcinoma provides the necessary parameters. METHODS: Sixty-nine BALB / C nude mice were synchronized in a programmed, independent light system (12 h light, 12 h dark) for at least 3 weeks. Then the bilateral axillary subcutaneous inoculation of human nasopharyngeal carcinoma cells (CNE-2) nude mice transplanted tumor, tumorigenesis at 6 different time points after the mice were killed in batches to collect bone marrow cells, fixed, stained flow Cytometry was used to measure the DNA content. The significance of the difference of cells at different time points was tested by ANOVA. Cosinor method was used to determine whether the distributions of cells in G1, S, G2 / M phase met the cosine function at 24h. Part of myeloid cells were lysed, and the protein levels of p53, p21 and Bcl-2 at different time points were detected by western blot. Results: The distributions of G1, S, G2 / M cells in 3,7,11,15,19,23HALO (Hours After Light Onset 3,7,11,19,23h) were significantly different with time . The change of 24h in G2 / M phase accorded with the cosine rhythm and the peak values were located at 10.8HALO and 1.8HALO respectively. Bone marrow cells p53 and Bcl-2 protein expression intensity 24h different. p53 was the strongest at 11HALO and the weakest at 7HALO; Bcl-2 was the strongest at 15HALO, the weakest at 19HALO, and p21 protein was not expressed. CONCLUSION: The DNA synthesis of human nasopharyngeal carcinoma nude mouse bone marrow cells changes regularly with the circadian rhythm. The expressions of p53 and Bcl-2 protein fluctuate regularly with the time of day and night.