揭示m6A甲基化在SLC22A3表达对乳腺癌干预中的保护作用。

Unraveling the protective role of m6A methylation in SLC22A3 expression for breast Cancer intervention.

作者信息

Xiao Yu, Cao Congcong, Zhang Yaqi, Zhao Chao, Liu Yuchen

机构信息

Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Breast and Thyroid Department, Shenzhen 518035, China.

Shenzhen Institute of Translational Medicine, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen 518035, China.

出版信息

Biochim Biophys Acta Mol Basis Dis. 2025 Oct;1871(7):167962. doi: 10.1016/j.bbadis.2025.167962. Epub 2025 Jun 24.

DOI:10.1016/j.bbadis.2025.167962

PMID:40570980

Abstract

BACKGROUND

The interplay between SLC22A3 expression and m6A RNA methylation is emerging as a significant factor in breast cancer pathology, yet the specific correlations and underlying mechanisms remain unclear.

METHODS

The prognostic significance of SLC22A3 expression in breast cancer was assessed using the Cancer Genome Atlas data. In vitro experiments were conducted using a custom-engineered dCas13b-METTL3 plasmid, designed to selectively enhance m6A methylation of SLC22A3 mRNA. Functional alterations in breast cancer cells post-transfection were evaluated. Additionally, overexpression of the m6A reader protein IGF2BP2 was achieved, and subsequent changes in SLC22A3 expression and overall breast cancer cell transcriptome were analyzed. MeRIP-seq and mRNA-seq were employed to analyze the expression and m6A methylation of Slc22a3 in a transgenic mouse model of breast cancer.

RESULTS

Bioinformatics analysis demonstrated a significant reduction in SLC22A3 expression in cancer tissues compared to adjacent non-cancerous tissues. Enhancement of SLC22A3 mRNA m6A methylation via the dCas13b-M3 plasmid in breast cancer cells led to increased SLC22A3 expression, accompanied by reduced cell proliferation and migration and induced apoptosis. Overexpression of IGF2BP2 similarly increased SLC22A3 expression. Further, RNA-seq identified 25 genes downstream of SLC22A3. Analysis of breast cancer tissues from mice revealed a decrease in both SLC22A3 expression and its m6A methylation as the breast cancer progressed.

CONCLUSION

SLC22A3 acts as protective factor in breast cancer. Enhanced m6A methylation of SLC22A3 mRNA and overexpression of the m6A reader IGF2BP2 upregulate its expression. The induction of SLC22A3 mRNA methylation through the m6A CRISPR approach effectively mitigates the malignancy of breast cancer cells.

摘要

背景

SLC22A3表达与m6A RNA甲基化之间的相互作用正逐渐成为乳腺癌病理学中的一个重要因素，但其具体相关性和潜在机制仍不清楚。

方法

利用癌症基因组图谱数据评估SLC22A3表达在乳腺癌中的预后意义。使用定制设计的dCas13b-METTL3质粒进行体外实验，该质粒旨在选择性增强SLC22A3 mRNA的m6A甲基化。评估转染后乳腺癌细胞的功能改变。此外，实现了m6A读取蛋白IGF2BP2的过表达，并分析了SLC22A3表达和整体乳腺癌细胞转录组的后续变化。采用MeRIP-seq和mRNA-seq分析乳腺癌转基因小鼠模型中Slc22a3的表达和m6A甲基化。

结果

生物信息学分析表明，与相邻的非癌组织相比，癌组织中SLC22A3表达显著降低。通过dCas13b-M3质粒增强乳腺癌细胞中SLC22A3 mRNA的m6A甲基化导致SLC22A3表达增加，同时细胞增殖和迁移减少并诱导凋亡。IGF2BP2的过表达同样增加了SLC22A3的表达。此外，RNA-seq鉴定出SLC22A3下游的25个基因。对小鼠乳腺癌组织的分析显示，随着乳腺癌的进展，SLC22A3表达及其m6A甲基化均降低。