Uncovering Metastasis Gene Expression Signatures and Unique Traits in Migrating Cancer Cells Using mRNA Sequencing and Transwell Assays
Yobin Kim
The Hotchkiss School, Lakeville, United States
Publication date: May 31, 2025
The Hotchkiss School, Lakeville, United States
Publication date: May 31, 2025
DOI: http://doi.org/10.34614/JIYRC2025I16
ABSTRACT
Understanding the molecular determinants that distinguish cells with high metastatic potential from their low-metastatic counterparts is critical for the development of improved diagnostic, prognostic, and therapeutic strategies. This study investigated the molecular characteristics of highly metastatic cancer cells selected via transwell assays from three human cell lines (A172, MCF7, and MDA-MB-231). This study aimed to determine specific gene expression profiles that define high metastatic and low metastatic cells. After three rounds of transwell selection, the cells that passed through the transwell membrane had elongated shape, indicating increased motility. The RNA-seq analysis identified differential gene expression signatures associated with metastasis, identifying 11 significantly upregulated genes and 209 downregulated genes. These changes in the expression of genes are involved in cell proliferation, biosynthesis, and genomic stability, as well as a decrease in immune response and antigen presentation. These changes indicate that the metastatic cells acquire a new environment, restructure their components, and avoid immune recognition to enhance invasive growth. By integrating phenotypic and transcriptomic data, this work highlights key molecular targets and pathways associated with metastasis. This result provides a foundation for developing more effective diagnostic markers and personalized therapeutic strategies.
Understanding the molecular determinants that distinguish cells with high metastatic potential from their low-metastatic counterparts is critical for the development of improved diagnostic, prognostic, and therapeutic strategies. This study investigated the molecular characteristics of highly metastatic cancer cells selected via transwell assays from three human cell lines (A172, MCF7, and MDA-MB-231). This study aimed to determine specific gene expression profiles that define high metastatic and low metastatic cells. After three rounds of transwell selection, the cells that passed through the transwell membrane had elongated shape, indicating increased motility. The RNA-seq analysis identified differential gene expression signatures associated with metastasis, identifying 11 significantly upregulated genes and 209 downregulated genes. These changes in the expression of genes are involved in cell proliferation, biosynthesis, and genomic stability, as well as a decrease in immune response and antigen presentation. These changes indicate that the metastatic cells acquire a new environment, restructure their components, and avoid immune recognition to enhance invasive growth. By integrating phenotypic and transcriptomic data, this work highlights key molecular targets and pathways associated with metastasis. This result provides a foundation for developing more effective diagnostic markers and personalized therapeutic strategies.
