Abstract：
Metastasis is an important hallmark of malignant tumors, and telomerase often exhibits high expression in these tumors. Monitoring the real-time dynamics of telomerase will provide valuable insights into its association with tumor metastasis. In this study, we described a microfluidic system for screening highly metastatic sublines based on differential cell invasiveness, investigated telomerase expression in the process of tumor metastasis and explored the genes and signaling pathways involved in tumor metastasis. Cells with different metastasis abilities were efficiently classified into different channels, and the fluorescence imaging visually demonstrates that cells with higher metastasis ability have stronger telomerase activity. In addition, we successfully established the high-metastasis-ability LoVo subline (named as LoVo-H) and low-metastasis-ability LoVo subline (named as LoVo-L) from the human colorectal cancer LoVo cell lines through only one round of selection using the system. The results show that the LoVo-H cells display superior proliferation and invasiveness compared to LoVo-L cells. Furthermore, 6776 differentially expressed genes of LoVo-H compared with LoVo-L were identified by transcriptome sequencing. The genes associated with telomerase activity, cell migration and the epithelial to mesenchymal transition were up-regulated in LoVo-H, and PI3K-Akt signaling pathway, extracellular matrix-receptor interaction and Rap1 signaling pathway were significantly enriched in LoVo-H. This microfluidic system is a highly effective tool for selecting highly metastatic sublines and the LoVo-H subline established through this system presents a promising model for tumor metastasis research. Furthermore, this work preliminarily reveals telomerase expression during tumor metastasis and provides a new strategy for studying tumor metastasis and cancer diagnosis.
摘要：
转移是恶性肿瘤的重要标志,和端粒酶通常在这些肿瘤中表现出高表达。监测端粒酶的实时动态将为其与肿瘤转移的关联提供有价值的见解。在这项研究中,我们描述了一种基于差异细胞侵袭性筛选高转移性亚系的微流体系统，研究了端粒酶在肿瘤转移过程中的表达，并探讨了参与肿瘤转移的基因和信号通路。具有不同转移能力的细胞被有效地分类到不同的通道，荧光成像显示转移能力较高的细胞具有较强的端粒酶活性。此外,我们仅通过使用该系统进行一轮筛选，就成功地从人类结直肠癌LoVo细胞系中建立了高转移能力LoVo亚系（命名为LoVo-H）和低转移能力LoVo亚系（命名为LoVo-L）。结果显示,与LoVo-L细胞相比,LoVo-H细胞显示出优异的增殖和侵袭性。此外，通过转录组测序鉴定了与LoVo-L相比的6776个差异表达的LoVo-H基因。与端粒酶活性相关的基因，LoVo-H中细胞迁移和上皮向间充质转化上调，和PI3K-Akt信号通路，LoVo-H显著富集了细胞外基质-受体相互作用和Rap1信号通路。该微流体系统是用于选择高转移性亚系的高效工具,并且通过该系统建立的LoVo-H亚系为肿瘤转移研究提供了有希望的模型。此外，这项工作初步揭示了肿瘤转移过程中端粒酶的表达，为研究肿瘤转移和癌症诊断提供了新的策略。
