Super-enhancer–driven EFNA1 fuels tumor progression in cervical cancer via the FOSL2-Src/AKT/STAT3 axis
Shu-Qiang Liu, … , Chun-Ling Luo, Jin-Xin Bei
Shu-Qiang Liu, … , Chun-Ling Luo, Jin-Xin Bei
Published February 18, 2025
Citation Information: J Clin Invest. 2025;135(8):e177599. https://doi.org/10.1172/JCI177599.
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Research Article Cell biology Oncology

Super-enhancer–driven EFNA1 fuels tumor progression in cervical cancer via the FOSL2-Src/AKT/STAT3 axis

Abstract

Super-enhancers (SEs) are expansive cis-regulatory elements known for amplifying oncogene expression across various cancers. However, their role in cervical cancer (CC), a remarkable global malignancy affecting women, remains underexplored. Here we applied integrated epigenomic and transcriptomic profiling to delineate the distinct SE landscape in CC by analyzing paired tumor and normal tissues. Our study identifies a tumor-specific SE at the EFNA1 locus that drives EFNA1 expression in CC. Mechanically, the EFNA1-SE region contains consensus sequences for the transcription factor FOSL2, whose knockdown markedly suppressed luciferase activity and diminished H3K27ac enrichment within the SE region. Functional analyses further underlined EFNA1’s oncogenic role in CC, linking its overexpression to poor patient outcomes. EFNA1 knockdown strikingly reduced CC cell proliferation, migration, and tumor growth. Moreover, EFNA1 cis-interacted with its receptor EphA2, leading to decreased EphA2 tyrosine phosphorylation and subsequent activation of the Src/AKT/STAT3 forward signaling pathway. Inhibition of this pathway with specific inhibitors substantially attenuated the tumorigenic capacity of EFNA1-overexpressing CC cells in both in vitro and in vivo models. Collectively, our study unveils the critical role of SEs in promoting tumor progression through the FOSL2-EFNA1-EphA2-Src/AKT/STAT3 axis, providing new prognostic and therapeutic avenues for CC patients.

Authors

Shu-Qiang Liu, Xi-Xi Cheng, Shuai He, Tao Xia, Yi-Qi Li, Wan Peng, Ya-Qing Zhou, Zi-Hao Xu, Mi-Si He, Yang Liu, Pan-Pan Wei, Song-Hua Yuan, Chang Liu, Shu-Lan Sun, Dong-Ling Zou, Min Zheng, Chun-Yan Lan, Chun-Ling Luo, Jin-Xin Bei

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Figure 2

Identification of EFNA1 as an SE-driven gene in CC.

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Identification of EFNA1 as an SE-driven gene in CC. (A) Schematic diagra...
(A) Schematic diagram illustrating H3K27ac ChIP-Seq signals proximate to the EFNA1 locus in SiHa and HCC-94 cells and the CRISPR/Cas9–mediated deletions targeting EFNA1-SEs. (B) Topologically associated domain (TAD) regions at the EFNA1 locus, predicted based on Hi-C data from CC cells. The heatmap color gradient, from white to red, represents the interaction intensity between the SE and the EFNA1 promoter region, ranging from low to high. (C and D) Analysis of EFNA1 expression in SiHa cells following EFNA1-SE deletions, including E1 (E1-KO), E2 (E2-KO), and promoter (P-KO) regions. mRNA levels were measured by quantitative reverse transcription–PCR (qRT-PCR) (C), and protein levels were assessed by Western blot (D), with β-actin serving as the internal control. (E) Dual-luciferase reporter assays in HEK293T cells assessing the enhancer activities of EFNA1 promoter (EFNA1-P) and EFNA1-SEs (EFNA1-E1-P, EFNA1-E2-P). (F and G) EFNA1 expression in CC cells treated with various concentrations of JQ1. qRT-PCR results (F) and Western blot results (G) are shown, with vehicle-treated cells as the control. (H) qPCR assay showing H3K27ac enrichments at the EFNA1 promoter (P) and SE regions (E1, E2) from ChIP assays in CC cells. The ChIP assays were conducted using H3K27ac antibodies, in cells treated either with or without 200 nM JQ1 for 24 hours. (I and J) Luciferase reporter assays assessing the transcriptional activity of the EFNA1 promoter (1.2 K and 2.5 K) (I) and the combined EFNA1 promoter and SE regions, EFNA1-P-SEs (J), in HEK293T cells treated with 200 nM JQ1 or vehicle control for 24 hours (EFNA1-E1-P, EFNA1-E2-P). Data are presented as mean ± SD, with n = 3 replicates. Between-group comparisons: 1-way ANOVA test. Significant P values: **P < 0.01, ***P < 0.001.