应用肿瘤相关寡核苷酸芯片筛选乳癌差异表达基因

Screening of differentially expressed genes in breast cancer by tumor-re lated oligochip

目的 :应用寡核苷酸芯片技术研究乳癌基因表达谱。方法 :根据文献获取目的基因 ,查询相应基因mRNA序列 ,通过计算机设计并合成探针 ,将探针点样于经化学修饰的载玻片上 ,制备含 2 88种基因的肿瘤相关基因寡核苷酸芯片。提取乳癌与相应正常乳腺组织总RNA ,通过逆转录制备荧光标记的cDNA探针并与芯片杂交 ,经洗片后扫描获取图像 ,计算机分析比较乳癌组织与正常乳腺组织的差异表达基因。结果 :检测 16例乳癌组织与正常乳腺组织标本 ,有 10例基因表达存在明显差异 ,其中表达增高的有 6种 ,表达降低的有 4种。结论 :乳癌组织与正常乳腺组织存在部分差异表达的基因 ,乳癌的发生发展与这些基因密切相关 ;寡核苷酸芯片技术在乳癌相关基因的研究中具有重要意义。

Objective: To study the gene expression p ro file of breast cancer using oligochip. Methods: Tumor asso ciated genes were obtained from related articles. Through Genbank, the mRNA seq u ence of the genes were acquired. The probes were designed by software according to the sequence and then synthesized. The synthesized probes were spotted on the chemical treated glass slide to fabricate oligochip containing 288 genes.Throug h reverse-transcription, first strand cDNA of breast cancer and normal bre a st tissues were synthesized and labeled with Cy3 or Cy5, respectively. After hy b ridization with the labeled samples and then washing, the oligochip was scanned and the acquired images were analyzed by software to find differentially express ed genes between the breast cancer and normal tissues. Results: The results of 16 samples showed that among the 288 genes in the chip,10 genes differentially expressed were identified.Expression of six of the 10 genes were up-regulated and that of the other 4 were down-regulated significantly. Conclusion: There are some differentially expressed genes between the breast cancer and normal tissues. The genes are related to the evolu tion of breast cancer. The application of oligochip is helpful in the investigat ion of the gene expression profiling of breast cancer.