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目的：探索荷人肺癌裸鼠上肿瘤细胞Bcl-2 表达及其活性生物时间节律差异。方法：体外培养A549 人肺癌细胞系，移植到裸小鼠身上，10 天成瘤后，随机分为6 大组，每大组再分2 小组，每组4 只，一组不给任何处理，设为对照，各组小鼠分别于光照后不同时间点取肿瘤细胞制成细胞悬液，固定染色后，用流式细胞仪以每个样品检测10000 个细胞的数量检测单个细胞的荧光强度，用流式细胞仪检测细胞周期情况，单因素方差分析法检验各期细胞在6 个时间点差异的显著性，用Cosinor 法考察G1，S，G2，M 期细胞在24h 的分布是否符合余弦函数，即是否有时辰节律。按6 个时间点取的肿瘤细胞，匀浆后裂解细胞，Western Blot 法测定 Bcl-2 的表达。结果：1. 结果发现肿瘤的生长曲线高峰出现在睡眠期中点，其次在活动期中点；G1、S、G2 期细胞变化符合余弦节律；2. Bcl-2 的表达在光照后7h 和19h 达到峰值，变化趋势与肿瘤细胞的周期性改变一致。结论：荷人肺癌裸鼠上肿瘤细胞的细胞周期可能随昼夜交替呈节律性变化，Bcl-2 的表达变化与肿瘤细胞的节律性改变一致。

英文摘要:

Objective: To explore the difference of the expression of Bcl-2 in tumor cells of nude mice bearing lung neoplasm and its activity circadian rhythm. Methods: Transplanted the A549 lung neoplasm cell lines which were cultured in vitro to nude mice. After tumorigenesis in 10 days, they were divided into 6 groups randomly. And then, every group was subdivided into 2 groups, with 4 mice each. One group served as control, and did not receive any treatments. As for other groups, the tumor cells of each group were made into cell suspension at different time points after irradiation. After fixing dye, 10000 cells in each sample were made out to detect single cell fluorescence intensity and the cell cycle by flow cytometry. Changes at six time-points of cells at each stage were detected by Single factor variance analysis test. The Cosinor method was used to investigate whether the distribution of cells in G1,S and G2, M phases during the 24h have a cosine function, that is, have an hour rhythm or not. Western Blot Determination was used to detect the Bcl-2 expression of tumor cells at six time-points after homogenate and lysis. Results: 1 The results showed that the tumor growth curve peaked in the sleep period mid-points, followed by mid-term point in the activity; cell changes at G1, S, and G2 stages are consistent with a cosine rhythm; 2 Bcl -2 expression reached peak at 7h and 19h after light, and changing trends were consistent with the cyclical nature of tumor cells. Conclusion: Tumor cells in nude mice bearing human lung cancer may have a cell cycle with rhythmic changes depending on day and night alternation. The expression of Bcl-2 changes with the rhythm of the tumor cells.

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