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Archive > Volume 31 Issue 8 > 2024,31(8):759-768. DOI:10.3872/j.issn.1007-385X.2024.08.003 Prev Next
Basic Research
Single-cell sequencing reveals the inhibitory role of Ly6E on DC infiltration and function in the microenvironment of 4T1 breast cancer cell transplanted tumor
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Abstract:
Objective: To preliminarily explore the potential mechanisms by which lymphocyte antigen 6 complex, locus E (Ly6E) regulates tumor immune microenvironment (TIME) in breast cancer 4T1 cells using single-cell RNA transcription sequencing technology (scRNA-seq). Methods: A mouse breast cancer 4T1 cell line with Ly6e gene knockout (Ly6E-KO) was constructed by CRISPR-Cas9 technology. The in vitro proliferation and in vivo growth capabilities of Ly6E-KO cells were compared to those of the wide type 4T1 cells (Ly6E-WT). CD45-positive cells from both types of tumor tissues were sorted by flow cytometry and analyzed using scRNA-seq. The sequencing results were first screened for differentially expressed gene profiles using the Seurat software package and annotated based on marker genes. Then, Cellchat and Monocle2 software were used to analyze the cell-to-cell interactions and the evolutionary trajectories of specific immune cells. Results: Compared with Ly6E-WT, Ly6E-KO cells showed no significant difference in in vitro proliferation capability but demonstrated significantly reduced in vivo growth capability (PConclusion: Ly6e gene knockout in tumor cells can enhance the anti-tumor immune response by increasing DC activation and infiltration in TIME.
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