我们前期创新性的构建了一种体外结直肠癌细胞3D集体侵袭及细胞分离模型，利用光转化后细胞荧光不同流式分选Leader、Follower及Central cells进行微量测序，筛选并验证出差异表达基因CCL5、GLUT1及PLOD2。在体外抑制CCL5/CCR5/PI3K/AKT通路能下调GLUT1及PLOD2的表达，并抑制Leader cells集体侵袭。本研究拟通过检测差异表达基因在体内外集体侵袭模型及人结直肠癌组织中的表达情况并利用差异表达基因分选Leader cells，进一步在Leader cells中验证CCL5/CCR5通过PI3K/AKT通路调控GLUT1及PLOD2的表达，从而分别增加Leader cells的糖酵解及周围基质硬度，领导肿瘤细胞团运动并促进集体侵袭。本项目旨在明确Leader cells在结直肠癌集体侵袭中的作用及相关分子机制，为结直肠癌治疗提供新的策略。

In our previous researches, we successfully constructed a 3D collective invasion and cell isolation model of colorectal cancer cells in vitro. Utilization of different flow cytometry-fluorescence could sort Leader, Follower and Central cells for microsequencing, screened and verified the differential expression genes CCL5, GLUT1 and PLOD2 as our target. Inhibition of CCL5/CCR5/PI3K/AKT pathway in vitro can down-regulate the expression of GLUT1 and PLOD2, moreover, inhibit the collective invasion of leader cells. This study intends to detect differential expression genes expression of collective invasion mode in vivo and vitro and human colorectal cancer tissues, and use differential expression genes to sort Leader cells. To further, we will verify that CCL5/CCR5 regulates GLUT1 and PLOD2 expression through the PI3K/AKT signaling pathway in Leader cells, and Leader cells lead the tumor cell mass to move and promote the collective invasion by increasing the self-glycolysis and hardness of the surrounding matrix. This project aims to clarify the role and molecular mechanism of Leader cell in collective invasion of colorectal cancer, and provide a new treatment strategy for colorectal cancer treatment.