Hu Qinghong, Zuo Xiaoxiao, Gu Xiaobin, Liu Liya, Tang Ying, Niu Xiaomin, Shi Yonggang, Han Liping
Department of Radiation Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450000, China.
Department of Gynecology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450000, China.
Sci Rep. 2025 Jul 1;15(1):20467. doi: 10.1038/s41598-025-05435-w.
Acquired radioresistance critically challenges cervical cancer radiotherapy management. Clinically relevant radioresistant cell models remain scarce, and CXCL8's role in cervical cancer-despite its tumorigenic/therapy-resistant associations in other cancers-is poorly characterized. Two radioresistant cervical cancer cell strains were established. mRNA-seq and bioinformatics analysis of radiosensitivity regulators identified CXCL8 as a key mediator. In vitro, assays of cell viability, clone formation, apoptosis and cell cycle were conducted following transient transfection of cervical cancer radiotherapy-resistant cell strains with knockdown of CXCL8, as well as subsequent addition of exogenous CXCL8 to cervical cancer parental cell strains. Radioresistant cervical cancer cell lines (Hela-RR/Siha-RR) were established through clinical protocol-mimicking irradiation, validated via proliferation/clonogenic/cell cycle assays. mRNA-seq identified 50 co-upregulated and 54 co-downregulated genes in resistant strains, with CXCL8 among top differentially expressed genes (IL11, CXCL8, MMP1, HSPA8, CA9, PPFIA4, EDN2, GUCY1A2, EFNA3, TNFAIP6). qRT-PCR confirmed CXCL8, TNFAIP6, SRNA8 and PPFIA4 dysregulation. Cox regression analysis of 96 candidate radiosensitivity regulators prioritized CXCL8 among eight key genes in cervical cancer. GEPIA2 and immunohistochemistry revealed CXCL8 overexpression in tumors. Functional studies demonstrated CXCL8 knockdown sensitized resistant cells to radiation, while exogenous CXCL8 induced resistance in parental lines.
获得性放射抗性对宫颈癌放疗管理构成了严峻挑战。临床上相关的放射抗性细胞模型仍然稀缺，并且尽管CXCL8在其他癌症中与肿瘤发生/治疗抗性相关联，但其在宫颈癌中的作用仍未得到充分表征。建立了两种放射抗性宫颈癌细胞系。通过对放射敏感性调节因子进行mRNA测序和生物信息学分析，确定CXCL8为关键介质。在体外，对宫颈癌放疗抗性细胞系进行CXCL8敲低的瞬时转染，随后向宫颈癌亲本细胞系中添加外源性CXCL8，之后进行细胞活力、克隆形成、凋亡和细胞周期检测。通过模拟临床方案的照射建立了放射抗性宫颈癌细胞系（Hela-RR/Siha-RR），并通过增殖/克隆形成/细胞周期检测进行了验证。mRNA测序确定了抗性菌株中50个共上调基因和54个共下调基因，CXCL8是差异表达最高的基因之一（IL11、CXCL8、MMP1、HSPA8、CA9、PPFIA4、EDN2、GUCY1A2、EFNA3、TNFAIP6）。qRT-PCR证实了CXCL8、TNFAIP6、SRNA8和PPFIA4的失调。对96个候选放射敏感性调节因子进行Cox回归分析，将CXCL8列为宫颈癌八个关键基因之一。GEPIA2和免疫组织化学显示肿瘤中CXCL8过表达。功能研究表明，CXCL8敲低使抗性细胞对辐射敏感，而外源性CXCL8诱导亲本细胞系产生抗性。
