人淋巴细胞来源的外泌体mRNAs作为辐射生物标志物的筛选与验证

doi:10.3969/j.issn.1004-616x.2026.01.007

Abstract

Abstract: OBJECTIVE： To investigate effects of ionizing radiation on the mRNA expression profile in exosomes from the AHH-1 human peripheral blood B lymphoblast cell line and to investigate usefulness of the profile as radiation biomarkers. METHODS： Using the AHH-1 cell line as a model， culture supernatants were collected from control cells and cells exposed to 2 Gy or 5 Gy of γ-rays at 24 h postirradiation. Exosomes were isolated by differential ultracentrifugation and characterized via nanoparticle tracking analysis for size distribution，transmission electron microscopy for morphology，and Western blotting for the surface markers CD63 and TSG101. Differentially expressed mRNAs were screened using expression microarray and subsequently subjected to GO and KEGG enrichment analyses. To further examine the dose- and time-dependent responses of candidate genes， AHH-1 cells were irradiated with 0， 1， 2， 4， or 6 Gy of γ-rays，and exosomes were collected at 24 h and 48 h postirradiation. Exosomal RNA was extracted，and the relative expression levels of selected mRNAs were validated by quantitative realtime PCR. RESULTS： Compared to the control group，a total of 197 differentially expressed mRNAs were identified in the exosomes from the 2 Gy irradiated cells，with 184 being upregulated and 13 downregulated. Following 5 Gy irradiation， 192 differentially expressed mRNAs were identified， comprising 170 upregulated and 22 downregulated transcripts. Bioinformatic analysis indicated that these altered mRNAs influenced cellular processes such as proliferation and apoptosis by participating in pathways including the TGF-β and ErbB signaling pathways. Real-time quantitative PCR analysis of the relative expression changes of candidate genes revealed that at 24 h after 0-6 Gy γ-rays irradiation， ZNFX1， DDB2， SPDYE2B， and CHD3 mRNA exhibited a significant upregulation trend. At 48 h after 4 Gy γ-rays irradiation， LENG8， FBXW7， AKAP1， and ZGRF1 mRNA were significantly upregulated. Notably，the expression levels of LENG8 and FBXW7 mRNA at 48 h after 4 Gy irradiation were approximately three times that of the control group. CONCLUSION： Ionizing radiation significantly altered mRNA expression profile in exosomes derived from AHH-1 cells. The differentially expressed genes identified (ZNFX1， DDB2， SPDYE2B， CHD3， LENG8， FBXW7， AKAP1， and ZGRF1) showed potential as biomarkers for ionizing radiation exposure.

Key words: AHH-1 cell line, human peripheral blood B lymphocyte, exosomes, mRNAs, radiation exposure, gene expression, biomarkers

CLC Number:

Q691.5

ZHOU Ruixia, LIU Jun, LU Xue, CAI Tianjing, YAN Juan, LIU Qingjie, LI Shuang. Preliminary validation of exosome mRNAs from human lymphocytes as radiation biomarkers[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2026, 38(1): 41-47,70.

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Preliminary validation of exosome mRNAs from human lymphocytes as radiation biomarkers

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