| 郭万峰,林汝仙,黄坚,郭国祯,王升启.辐射相关基因芯片的制备及其在肺癌细胞中的应用[J].中华放射医学与防护杂志,2005,25(2):107-110 |
| 辐射相关基因芯片的制备及其在肺癌细胞中的应用 |
| Preparation of oligonucleotide microarray for radiation-associated gene expression detection and its application in lung cancer cell lines. |
| 投稿时间：2004-10-25 |
| DOI： |
| 中文关键词: 基因芯片 电离辐射 肺癌细胞系 |
| 英文关键词:Microarray Ionizing radiation lung cancer cell lines |
| 基金项目: |
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| 中文摘要: |
| 目的 寻找参与肺癌细胞辐射抗性的基因。方法 构建辐射相关的基因芯片,初步筛选出在不同辐射敏感性肺癌细胞系中差异表达的基因。为了评价芯片结果的可靠性,我们对一些差异表达基因进行了RTPCR验证。结果 和辐射敏感的NCIH446细胞相比,辐射抗性的A549中有18个基因有明显的改变,8个基因是上调,10个基因是下调。在照射后6h和24h,A549细胞中分别有22个基因(19个基因上调,3个下调)和26个基因(8个上调,18个下调)的差异表达;NCIH446细胞中分别有17个基因(9个基因上调,8个下调)和18个基因(6个上调,12个下调)差异表达。从这些基因中,我们发现一些参与细胞增殖和抗凋亡的基因,在照射后的A549细胞中是增高的,而在NCIH446细胞中是降低的。另外一些参与DNA修复的基因,在A549中比在NCIH446细胞中有更高的表达。结论 一些参与细胞DNA修复、细胞周期调控、细胞增殖和抗凋亡作用的基因可能有助于A549细胞辐射抗性的形成。 |
| 英文摘要: |
| Objective The response of tumor cell to radiation is accompanied by complex change in patterns of gene expression. It is highly probable that a better understanding of molecular and genetic changes can help to sensitize the radioresistant tumor cells. Methods Oligonucleotide microarray provides a powerful tool for high-throughput identifying a wider range of genes involved in the radioresistance. Therefore, we designed one oligonucleotide microarray according to the biological effect of IR. By using different radiosensitive lung cancer cell lines, we identified genes showing altered expression in lung cancer cell lines. To provide independent confirmation of microarray data, semi-quantitative RT-PCR was performed on a selection of genes. Results In radioresistant A549 cell lines, a total of 18 genes were selected as having significant fold-changes compared to NCI-H446, 8 genes were up-regulated and 10 genes were down-regulated. Subsequantly, A549 and NCI-H446 cells were delivered by ionizing radiation. In A549 cell line, we found 22 (19 up-regulated and 3 down-regulated) and 26 (8 up-regulated and 18 down-regulated) differentially expressed genes at 6h and 24h after ionizing radiation. In NCI-H446 cell line, we identified 17 (9 up-regulated and 8 down-regulated) and 18 (6 up-regulated and 12 down-regulated) differentially expressed genes at 6 h and 24 h after ionizing radiation. We tested seven genes (MDM2, p53,, XRCC5, Bcl-2, PIM2, NFKBIA and Cyclin B1) for RT-PCR, and found that the results were in good agreement with those from the microarray data except for NFKBIA gene, even though the value for each mRNA level might be different between the two measurements. In present study, we identified some genes with cell proliferation and anti-apoptosis, such as MDM2, BCL-2, PKCz and PIM2 expression levels increased in A549 cells and decreased in NCI-H446 cells after radiation, and other genes with DNA repair, such as XRCC5, ERCC5, ERCC1, RAD9A, ERCC4 and DNA-PK activities were higher in A549 cells than NCI-H446 cells. Conclusion These results support a few genes with DNA repair, regulation of cell cycle, cell proliferation and anti-apoptosis as candidates for radioresistance of A549 lung cancer cells. These maybe, provide several gene targets to sensitize the radioresistant cells for improving the radiocurability of lung cancer. |
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