胆管癌放射敏感相关基因的基因芯片实验研究

Differentially Expressed Radiosensitivity-associated Genes in Cholangiocarcinoma Detected by cDNA Microarray

目的:应用cDNA芯片技术进行肝外胆管癌放射敏感性相关基因表达谱差异分析。方法:将10只荷人胆管癌的裸鼠移植瘤模型随机分为对照组和照射纽,分别给予10Gy、20Gy单次照射剂量,6h和24h后提取肿瘤组织,利用一步法提取组织块或细胞中的总RNA,浓缩、纯化,逆转录合成掺入荧光分子的cDNA链探针,与21 522条人PCR微矩阵芯片杂交,用双通道激光扫描仪进行扫描并分析表达谱的差异。结果:人胆管癌细胞QBC939在10Gy、20Gy单次照射剂量照射后的基因表达谱分析,发现306条基因表达差异,其中27条功能基因与剂量、照射后时间一致相关(14条表达上调、13条表达下调)。结论:基因芯片能快速筛选胆管癌放射敏感性相关基因,为提高放射治疗疗效提供生物学依据。

Objective： To search for differentially expressed radiosensitivity-associated genes in cholangiocarcinoma cell line QBC939 using cDNA microarrays. Methods： Mice bearing chalongiocarcinoma tumors were exposed to radiation （10Gy and 20Gy）. A total of 21,522 human genes were spotted on a kind of chemi- cal material coated glass slide. Total RNA was isolated from chalongiocarcinoma tissues in tumor-bearing mice and purified. The mRNA was reverse transcribed to cDNA with fluorescent dUTP in the reaction mixture to prepare hybridization probes that were then hybridized to the cDNA microarray. The fluorescent signals on the cDNA microarray were scanned using ScanArray Express. Differential expression was analyzed. Results： The gene expression profiles revealed 306 genes differentially expressed in cholangiocarcinoma samples after radiation, including 27 functional genes. The expression of 14 genes was upregulated and that of 13 genes was downregulated. Conclusion： Differentially expressed radiosensitivity-associated genes in cholangiocarcinoma can be screened quickly by cDNA microarray.