Research Center for Analytical Sciences, Northeastern University, Shenyang, 110819, PR China.
Research Center for Analytical Sciences, Northeastern University, Shenyang, 110819, PR China.
Talanta. 2024 Dec 1;280:126690. doi: 10.1016/j.talanta.2024.126690. Epub 2024 Aug 8.
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.
转移是恶性肿瘤的一个重要标志，端粒酶在这些肿瘤中常表现出高表达。监测端粒酶的实时动态将为研究其与肿瘤转移的关系提供有价值的见解。在这项研究中，我们描述了一种基于细胞侵袭性差异筛选高转移性亚系的微流控系统，研究了肿瘤转移过程中端粒酶的表达，并探讨了涉及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中，荧光成像直观地表明，具有更高转移能力的细胞具有更强的端粒酶活性。此外，我们成功地从人结直肠癌细胞系 LoVo 中通过该系统仅一轮选择就建立了高转移性能力的 LoVo 亚系（命名为 LoVo-H）和低转移性能力的 LoVo 亚系（命名为 LoVo-L）。结果表明，与 LoVo-L 相比，LoVo-H 细胞具有更强的增殖和侵袭能力。此外，通过转录组测序，我们鉴定了 LoVo-H 与 LoVo-L 相比有 6776 个差异表达基因。与端粒酶活性、细胞迁移和上皮间质转化相关的基因在 LoVo-H 中上调，PI3K-Akt 信号通路、细胞外基质-受体相互作用和 Rap1 信号通路在 LoVo-H 中显著富集。该微流控系统是一种高效的选择高转移性亚系的工具，通过该系统建立的 LoVo-H 亚系为肿瘤转移研究提供了一个很有前途的模型。此外，这项工作初步揭示了肿瘤转移过程中端粒酶的表达，并为研究肿瘤转移和癌症诊断提供了新的策略。
