Erianin reverses 5-FU resistance by targeting CALM1/CAMKK2 and activating autophagy in colorectal cancer

IF 5.4 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions Pub Date : 2025-09-19 DOI:10.1016/j.cbi.2025.111750

Fangyuan Zhou , Luorui Shang , Jinxiao Li , Mengqi Zhang , Shuhan Wang , Yuju Cai , Qifeng Lin , Haiyang Gao , Shenglan Yang

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引用次数: 0

Abstract

Background

Chemoresistance represents a significant factor contributing to the failure of clinical treatments in colorectal cancer (CRC), with resistance to 5-fluorouracil (5-FU) being notably prevalent. Erianin has demonstrated potential in reversing tumor resistance; however, its specific effects and underlying mechanisms in the context of 5-FU-resistant CRC require comprehensive validation.

Methods

The half-maximal inhibitory concentration (IC50) of Erianin and 5-FU was determined using the CCK8 assay. CRC cells resistant to 5-FU were induced by progressively increasing concentrations of 5-FU. Cellular proliferation, migration, and invasion were evaluated via 5-ethynyl-2′-deoxyuridine (EdU) assays, wound-healing assays, and Transwell Matrigel invasion assays. The expression levels of protein and mRNA expression levels for various molecules were quantified using Western blotting (WB) and quantitative real-time polymerase chain reaction (qRT-PCR). Immunofluorescence assays were employed to assess the expression of autophagy-related markers. The in vivo therapeutic efficacy of Erianin was assessed using a xenograft tumor model.

Results

Erianin effectively reinstated the sensitivity of 5-FU-resistant CRC cells to 5-FU, significantly reducing tumor cell proliferation, migration, and invasion, as well as inhibiting xenograft tumor growth. Mechanistic investigations demonstrated that Erianin targets and binds to the calmodulin 1 (CALM1) protein, enhancing its stability and subsequently increasing the phosphorylation levels of CAMKK2. This interaction facilitates autophagy in 5-FU-resistant CRC cells, ultimately leading to tumor cell death.

Conclusion

Erianin sensitized the resistant CRC cells to 5-FU by activating the CALM1/CAMKK2 signaling pathway, thereby inducing autophagy. The combination of 5-FU and Erianin presents a promising therapeutic approach to enhance survival outcomes in patients with refractory CRC.

Abstract Image

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Erianin通过靶向CALM1/CAMKK2和激活结直肠癌自噬逆转5-FU耐药

化疗耐药是导致结直肠癌（CRC）临床治疗失败的一个重要因素，其中5-氟尿嘧啶（5-FU）耐药尤为普遍。Erianin已被证明具有逆转肿瘤耐药的潜力；然而，其在5- fu耐药CRC中的具体作用和潜在机制需要全面验证。方法采用CCK8法测定苦参素和5-FU的半最大抑制浓度（IC50）。逐渐增加的5-FU浓度诱导CRC细胞对5-FU产生抗性。通过5-乙基-2 ' -脱氧尿苷（EdU）测定、伤口愈合测定和Transwell Matrigel侵袭测定来评估细胞增殖、迁移和侵袭。采用Western blotting （WB）和定量实时聚合酶链式反应（qRT-PCR）检测各组蛋白表达水平和各分子mRNA表达水平。免疫荧光法检测自噬相关标志物的表达。采用异种移植物肿瘤模型评价苦连宁的体内治疗效果。结果seranin能有效恢复5-FU耐药CRC细胞对5-FU的敏感性，显著降低肿瘤细胞的增殖、迁移和侵袭，抑制异种移植肿瘤的生长。机制研究表明，Erianin靶向并结合钙调蛋白1 (calmodulin 1, CALM1)，增强其稳定性，随后增加CAMKK2的磷酸化水平。这种相互作用促进了5- fu耐药CRC细胞的自噬，最终导致肿瘤细胞死亡。结论erianin通过激活CALM1/CAMKK2信号通路使CRC耐药细胞对5-FU致敏，从而诱导自噬。5-FU联合Erianin是提高难治性结直肠癌患者生存率的一种有希望的治疗方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Chemico-Biological Interactions 生物-毒理学

CiteScore

7.70

自引率

3.90%

发文量

410

审稿时长

36 days

期刊介绍： Chemico-Biological Interactions publishes research reports and review articles that examine the molecular, cellular, and/or biochemical basis of toxicologically relevant outcomes. Special emphasis is placed on toxicological mechanisms associated with interactions between chemicals and biological systems. Outcomes may include all traditional endpoints caused by synthetic or naturally occurring chemicals, both in vivo and in vitro. Endpoints of interest include, but are not limited to carcinogenesis, mutagenesis, respiratory toxicology, neurotoxicology, reproductive and developmental toxicology, and immunotoxicology.