随着CellSearch系统被FDA批准进入临床，CTC检测已成为乳腺癌预后判断、疗效监测的重要指标，对CTC基因表达的研究也已成为乳腺癌个体化医疗的新热点；但由于其自身限制，尚未有针对CTC基因表达检测技术的临床应用。课题组在传统ISET基础上，建立了改良ISET技术体系，可应用于乳腺癌CTC的检测；获取的CTC表面各种抗原或分子标记均无破坏，可供进一步分子生物学检测。.我们拟在此基础上，将含有乳腺癌CTC的滤膜进行细胞蜡块包埋，切片后进行免疫组化、FISH检测CTC相关基因（PR、ER、Her-2）的表达情况，建立CTC相关基因表达的检测方法；使用MCF-7细胞株和MCF-7细胞人乳腺癌异种移植裸鼠模型验证其有效性；同时针对10例转移性乳腺癌患者，检测其CTC相关基因表达情况，比较原发瘤、CTC及转移瘤相关基因表达的差异性，探讨其临床意义，进而为乳腺癌个体化治疗方案的制定提供新思路。

Circulating tumor cell (CTC) detection has become an important indicator for prognostic evaluation and therapeutic monitoring of breast cancer since the clinical utility of the CellSearch system is approved by the FDA. As a result, the gene expression of the CTC is highlighted for the individualized treatment of breast cancer. However, there is still lack of CTC gene expression detecting technologies which can be clinically used due to the limitations of the CellSearch system. In order to solve this problem, we improved the isolation by size of epithelial tumor cells（ISET）system and established a modified one. This new system is able to not only detect CTC of breast cancer patients, but also to intactly catch the antigens and molecular markers on the surface of CTC without any damage, thus is ideal for the biological analysis of CTC. .Based on this modified ISET system, we plan to collect CTC from breast cancer patients, embed the CTC as well as the filter membrane with paraffin, and then detect the expression of CTC-related genes (ER, PR, HER2) via immunohistochemistry and fluorescence in situ hybridization (FISH) analysis. In this way, we can build a detecting approach for these genes. We furtherly use breast cancer cell line MCF-7 and MCF-7 human breast cancer xenograft model in nude mice to verify the reliability of this approach. In addition, we will collect CTC from 10 patients with metastatic breast cancer and detect the expression of CTC-related genes. By comparing the gene expression differences among the primary tumor, CTC and metastatic tumor, it is expected to reveal the clinical value of the modified ISET, so as to provide new ideas to make the individualized treatment scheme of breast cancer.

随着CellSearch系统被FDA批准进入临床，CTC检测已成为乳腺癌预后判断、疗效监测的重要指标，对CTC基因表达的研究也已成为乳腺癌个体化医疗的新热点；但由于其自身限制，尚未有针对CTC基因表达检测技术的临床应用。课题组在传统ISET基础上，建立了改良ISET技术体系，可应用于乳腺癌CTC的检测；获取的CTC表面各种抗原或分子标记均无破坏，可供进一步分子生物学检测。我们在此基础上，将含有乳腺癌CTC的滤膜进行细胞蜡块包埋，切片后进行免疫组化、FISH检测CTC相关基因（PR、ER、Her-2）的表达情况，建立CTC相关基因表达的检测方法；使用MCF-7细胞株和MCF-7细胞人乳腺癌异种移植裸鼠模型验证了其切实可行；同时针对10例转移性乳腺癌患者，检测其CTC相关基因表达情况，并比较原发瘤、CTC及转移瘤相关基因表达的差异性，研究结果表明通过检测乳腺癌患者CTC相关基因的表达可应用于指导乳腺癌个体化治疗方案的制定，其相关于传统的病理活检具有创伤小、花费少、可实时性检测等优点。