| GUO Wan-feng,LIN Ru-xian,HUANG Jian.Preparation of oligonucleotide microarray for radiation-associated gene expression detection and its application in lung cancer cell lines.[J].Chinese Journal of Radiological Medicine and Protection,2005,25(2):107-110 |
| Preparation of oligonucleotide microarray for radiation-associated gene expression detection and its application in lung cancer cell lines. |
| Received:October 25, 2004 |
| DOI： |
| KeyWords:Microarray Ionizing radiation lung cancer cell lines |
| FundProject: |
| Author Name | Affiliation | E-mail | | GUO Wan-feng | Institute of Radiation Medicine Science, Academy of Military Medical Sciences, Beijing 100850, China | sqwang@nic.bmi.ac.cn | | LIN Ru-xian | Institute of Radiation Medicine Science, Academy of Military Medical Sciences, Beijing 100850, China | | | HUANG Jian | Institute of Radiation Medicine Science, Academy of Military Medical Sciences, Beijing 100850, China | | | 郭国祯 | 第四军医大学放射医学教研室 | | | 王升启 | Institute of Radiation Medicine Science, Academy of Military Medical Sciences, Beijing 100850, China | |
|
| Hits: 3263 |
| Download times: 2295 |
| Abstract:: |
| 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. |
| HTML View Full Text View/Add Comment Download reader |
| Close |
|
|
|
