细胞周期基因芯片筛选不同增殖状态下CNE-2细胞的差异表达基因

Differential Expressions of Cell Cycle Genes in Different Proliferation Status of CNE-2 Cell Lines

目的:了解不同增殖状态鼻咽癌细胞(CNE-2)细胞周期基因表达差异,探讨鼻咽癌放射治疗中加速再增殖的机理。方法:对不同增殖状态的CNE-2细胞,采用基因芯片技术、实时荧光定量PCR检测其细胞周期相关基因表达差异。结果:细胞增殖活性最高时相(2Gy连续照射第3天)与最低时相(2Gy连续照射第5天)表达有差异的基因有3条,为检测基因总数的3%(3/100),其中细胞增殖活性增高时上调基因为CCNE1、CUL-5,下调基因为CDKN1A。应用相对定量荧光实时PCR技术对部分差异表达在2倍以上的基因(CCNE1、CDKN1A))进行了mRNA水平的验证,结果显示实时PCR实验结果与芯片结果具有良好的一致性。结论:某些细胞周期基因如CC-NE1、CUL-5、CDKN1A等可能参与了鼻咽癌放射治疗过程中发生的加速再增殖过程,有必要对这些基因功能作进一步研究,以探明照射过程中肿瘤细胞加速再增殖发生的分子机制。

Objective： To understand the differentially gene expressions of nasopharyngeal carcinoma（NPC） CNE-2 cell lines in different proliferative status and to investigate the mechanism of accelerated repopulation in NPC during irradiation. Methods： In different proliferation status of CNE-2 cell lines, the differential gene expressions of cell cycle genes were detected by gene chip technology and real-time fluorescence quantitative PCR. Results： Between the peak phase （the 3rd day of continuous irradiated by a daily fraction of 2Gy）and the valley phase（ the 5th day of continuous irradiated by a daily fraction of 2Gy）of CNE-2 cell proliferative activity, 3 cell cycle genes were differentially expressed, accounting for 3% （3/100） of the detected genes. In the peak phase of cell proliferative activity, up-regulated genes were CCNE1 and CUL-5,down-regulated gene was CDKN1A. We analyzed several interesting differential genes which were up or down at least 2 fold in mRNA level （ CCNE1 ,CDKN1A） with the real-time fluorescence quantitative PCR. The analysis revealed a similar expression pattern to the microarray results. Conclusion ： The cell cycle genes such as CCNE1, CUL-5 and CDKN1A etc may take part in the accelerated repopulation process of CNE-2 cell lines during irradiation. The accelerated repopulation process may be a complicated and multifactorial process. Further study of these accelerated repopulation genes will provide the insights into the molecular mechanism of the accelerated repopulation process during irradiation.