乳腺癌miRNAs表达谱检测及初步分析

Microarray-based miRNAs profiling in human breast cancer

目的制作哺乳动物micro RNAs(miRNAs)检测微阵列,筛选乳腺癌miRNAs表达谱并进行初步分析。方法利用已知的人及小鼠的miRNAs序列,设计了496条探针,制作miRNAs微阵列并验证其可靠性;培养乳腺癌细胞株MCF-7和正常乳腺上皮细胞株MCF-10A,分别抽提细胞总RNA,分离小分子RNA,经T4RNA连接酶荧光标记后与微阵列杂交,通过芯片扫描和数据分析,获得乳腺癌miRNAs表达谱。结果成功制作哺乳动物miRNAs微阵列,筛选获得57个乳腺癌相关miRNAs,其中,29个为低表达,28个为高表达。结论建立了哺乳动物miRNAs检测微阵列的技术平台,并筛选到乳腺癌miRNAs表达谱,为进一步研究其在乳腺癌发病机制中的作用打下基础。

Objective To make microRNAs （miRNAs） microarray for mammal miRNAs profiling and to screen breast cancer-specific miRNAs and analyze their functions preliminarily. Methods First, 496 probes designed according to known miRNAs sequences of human and mouse. Then, miRNAs microarray was made and subjected to credibility verification. Next, MCF-7, breast cancer cell line, and MCF-10A, normal human mammary epithelial cell line were cultured for respective total RNA extraction. Small molecular RNA was separated and fluorescence labeling was performed by W4 RNA ligase. Finally, microarray hybridization was performed to obtain miRNAs signature of human breast cancer. Results We successfully made credible miRNAs microarray and obtained 57 breast cancer-related miRNAs by microarray-based screening, including 29 highlyexpressed miRNAs and 28 lowly-expressed ones. Conclusion We built up the technical platform of mammal miRNAs detection and obtained miRNAs signature of human breast cancer so as to lay necessary basis for research on the function of miRNAs in breast cancer.