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中国癌症防治杂志

中国癌症防治杂志 ›› 2025, Vol. 17 ›› Issue (3): 341-348.doi: 10.3969/j.issn.1674-5671.2025.03.12

• 其他消化系统肿瘤专栏 • 上一篇    下一篇

EGLN1介导自噬调控结直肠癌恶性表型的机制研究

  

  1. 广西医科大学附属肿瘤医院胃肠外科结直肠肛门病区；广西结直肠癌基础与转化研究重点实验室；广西壮族自治区卫生健康委员会结直肠癌防治重点实验室
  • 出版日期:2025-06-25 发布日期:2025-07-10
  • 通讯作者: 罗涛 E-mail:luotao@gxmu.edu.cn； 韦传毅 E-mail:weichuanyi@sr.gxmu.edu.cn
  • 基金资助:
    广西壮族自治区大学生创新创业训练计划（S202410598217）

Mechanistic study of EGLN1-mediated autophagy regulation in the malignant phenotypes of colorectal cancer

  • Online:2025-06-25 Published:2025-07-10

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摘要： 目的 探讨Egl⁃9家族缺氧诱导因子1（Egl⁃9 family hypoxia inducible factor 1，EGLN1）缺失对结直肠癌（colorectal cancer,CRC）细胞增殖、迁移、侵袭及自噬通路的影响，为CRC精准治疗提供潜在靶点。方法 利用CRISPR⁃Cas9技术构建EGLN1敲除的LoVo与RKO细胞株；通过CCK⁃8法、集落形成、Transwell实验及划痕愈合实验，分别检测细胞的增殖、侵袭及迁移能力。Western blot分析自噬标志物LC3B⁃Ⅱ/LC3B⁃Ⅰ值与p62蛋白表达水平，评估自噬通路激活情况。通过建立裸鼠皮下移植瘤模型，评估自噬激活剂雷帕霉素对EGLN1敲除异种移植瘤生长的影响。采用苏木精-伊红（hematoxylin and eosin，HE）染色和Ki⁃67免疫组化检测异种移植瘤组织细胞增殖情况。结果 成功构建稳定的EGLN1敲除细胞系。EGLN1的缺失显著增强LoVo和RKO细胞增殖、迁移和侵袭（均P<0.05）；自噬相关蛋白分析结果显示，EGLN1敲除后LC3B⁃Ⅱ/LC3B⁃Ⅰ值下降，p62显著积累。采用晚期自噬抑制剂氯喹或早期抑制剂3⁃甲基腺嘌呤处理进一步加剧自噬抑制。体内实验表明，EGLN1敲除组的肿瘤体积显著大于EGLN1野生型组（P<0.05），而雷帕霉素治疗有效抑制了肿瘤生长。HE染色和Ki⁃67免疫组化证实，EGLN1敲除促进异种移植瘤中的细胞增殖，而雷帕霉素可逆转该促瘤效应。结论 EGLN1通过激活自噬通路抑制CRC细胞的恶性表型，其缺失可能导致自噬抑制，从而促进肿瘤细胞增殖、迁移与侵袭。靶向EGLN1/自噬轴可能是治疗CRC的有效策略。

关键词: 结直肠癌, Egl-9家族缺氧诱导因子1, 增殖, 迁移, 侵袭, 自噬

Abstract: Objective To investigate the impact of Egl⁃9 family hypoxia⁃inducible factor 1 (EGLN1) deficiency on the proliferation, migration, invasion, and autophagic pathway of colorectal cancer (CRC) cells, and to provide potential targets for precision therapy in CRC. Methods EGLN1⁃knockout LoVo and RKO cell lines were generated using CRISPR⁃Cas9 technology. Cell proliferation, invasion, and migration were assessed through CCK⁃8 assay, colony formation assay, Transwell assay, and scratch wound⁃healing assay, respectively. Western blot analysis was performed to determine the LC3B⁃Ⅱ/LC3B⁃Ⅰratio and p62 protein expression levels, thereby assessing the activation of the autophagy pathway. A subcutaneous xenograft model in nude mice was developed to evaluate the effects of the autophagy activator Rapamycin on tumor growth in EGLN1 knockout xenografts. The cell proliferation of xenograft tissues was detected by hematoxylin and eosin (HE) staining and Ki⁃67 immunohistochemical staining. Results Stable EGLN1 knockout cell lines were successfully developed. The deletion of EGLN1 significantly enhanced proliferation, migration, and invasion in both LoVo and RKO cell lines (all P<0.05). The analysis results of autophagy⁃related proteins revealed a decreased LC3B⁃Ⅱ/LC3B⁃Ⅰratio and significant accumulation of p62 following EGLN1 knockout.  Treatment with the late⁃stage autophagy inhibitor chloroquine or the early⁃stage inhibitor 3⁃methyladenine  further exacerbated the inhibition of autophagy. In vivo experiments demonstrated that the EGLN1 knockout group developed significantly larger tumor volumes compared to the EGLN1 wild⁃type group (P<0.05), whlie treatment with Rapamycin effectively suppressed tumor growth. HE staining and Ki⁃67 immunohistochemistry confirmed that EGLN1 knockout promoted cell proliferation in xenografts, an effect that was reversible with rapamycin treatment. Conclusions EGLN1 appears to suppress the malignant phenotype of CRC cells by activating the autophagic pathway. Its deficiency may lead to  autophagy inhibition, thereby promoting tumor cell proliferation, migration, and invasion. Targeting the EGLN1/autophagy axis may represent an effective therapeutic strategy for CRC.

Key words: Colorectal cancer, Egl-9 family hypoxia inducible factor 1,  Proliferation, Migration,  Invasion, Autophagy

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