Leader cells通过CCL5调控糖酵解及基质硬度促进结直肠癌集体侵袭的 作用机制

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.

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。

许多实体肿瘤在集体侵袭过程中依赖Leader cells和Follower cells的调节和组织，但Leader cells在结直肠癌(CRC)集体侵袭中的特异性生物标志物和机制尚不清楚。本研究旨在鉴定Leader cells的特异性生物标志物，并揭示其在结直肠癌集体侵袭转移过程中的分子机制。通过在体外构建了3D光转换结直肠癌多细胞球模型，分离出Leader cells和Follower cells，RNA-Seq、功能和动物实验表明，Leader cells中的GLUT1、PLOD2和CCL5是CRC集体侵袭所必需的，CCL5在前导细胞中通过PI3K/Akt信号通路上调GLUT1和PLOD2的表达，此外，GLUT1和CCL5可作为CRC集体侵袭中先导细胞的特异性生物标志物，其共表达与CRC患者预后不良相关，值得注意的是，阻断GLUT1和CCL5能有效抑制CRC的集体侵袭。我们的研究结果表明，CCL5和GLUT1可能作为Leader cells的特异性标志物，并且作为缺氧诱导的代谢转移和胶原沉积的潜在关键调节因子，是结直肠癌集体入侵是必需的。