ORIGINAL RESEARCH article

Front. Immunol.

Sec. Cancer Immunity and Immunotherapy

Hsa-miR-99a deficiency contributes to MSI-H colorectal cancer progression by activating the mTOR pathway and inducing Th1/Th2 imbalance

    XY

    Xuejing Yang

    TZ

    Tingting Zhang

    HS

    Hu Sun

    HF

    Huijing Feng

    DS

    Dong Song

  • Shanxi Bethune Hospital, Shanxi Medical University, Taiyuan, China

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Abstract

Background: Hsa-miR-99a has been linked to the advancement of several malignancies, including colorectal cancer (CRC). This investigation seeks to elucidate its function and regulatory network in CRC. Methods: Differential expression of hsa-miR-99a was analyzed between microsatellite stable (MSS) and microsatellite instability-high (MSI-H) subgroups. Potential target mRNAs of hsa-miR-99a were retrieved from TargetScan and miRWalk databases. Overlapping mRNAs were subjected to correlation analysis, with candidate genes selected based on |correlation coefficient| > 0.3 and p < 0.05. Enrichment analyses were performed, highlighting key pathways, particularly mTOR signaling. Immune infiltration profiles were compared between MSS and MSI-H groups. Correlation between hsa-miR-99a and mTOR pathway-related genes was validated by RT-qPCR and Western blot. Additionally, multiplex immunohistochemistry (mIHC) with tyramine signal amplification (TSA) was used to characterize the immune microenvironment in MSI-H CRC samples. Results: Hsa-miR-99a expression was significantly higher in MSS compared to MSI-H groups. Twelve key target genes were identified, including ATP2B2, ATP2B4, SLC8A1, KCNJ5, NTRK3, TNFRSF19, GHR, CXCL12, NTNG1, SDC2, SFRP1, and PRICKLE2. Immune infiltration analysis revealed significant differences in 24 cell types between MSS and MSI-H groups, including Th1, Th2, Th17, and effector memory CD8+ T cells. Ten mTOR pathway-related genes (ATP6V1G2, WNT2, WNT6, WNT9A, WNT9B, FZD1, FZD4, FZD8, IGF1, AKT3) exhibited strong correlation with hsa-miR-99a. Among these, ATP6V1G2 and WNT6 were upregulated in the MSI-H group. mIHC analysis indicated reduced Th1 but increased Th2 and Th17 biomarkers in MSI-H CRC. Conclusion: This study identified key genes and immune microenvironment alterations regulated by hsa-miR-99a in CRC, offering novel insights and potential therapeutic targets for CRC treatment.

Summary

Keywords

colorectal cancer, hsa-miR-99a, Immuneinfiltration, Microsatellite Instability, mTOR pathway

Received

26 January 2026

Accepted

20 February 2026

Copyright

© 2026 Yang, Zhang, Sun, Feng and Song. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

*Correspondence: Dong Song

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