Transcription factor PRRX1-activated ANXA6 facilitates EGFR-PKCα complex formation and enhances cisplatin sensitivity in bladder cancer

IF 5.2 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Life sciences Pub Date : 2024-11-09 DOI:10.1016/j.lfs.2024.123228

Jinlong Cao , Siyu Chen , Jirong Wang , Xinpeng Fan , Shanhui Liu , Xiaoran Li , Li Yang

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Abstract

Background

Tumor resistance to cisplatin represents a major clinical challenge, particularly in bladder cancer (BC). ANXA6 is a member of annexin family, and its role in cisplatin resistance remains unclear. This study explores ANXA6's role in promoting cisplatin sensitivity.

Methods

Bioinformatics analyses and clinical specimen verifications assessed the correlation between ANXA6 and cisplatin treatment. A series of assays, including CCK-8, clone formation assay, flow cytometry assays for reactive oxygen species (ROS) and apoptosis, and comet assays, were used to confirm ANXA6's role in enhancing cisplatin sensitivity and re-sensitizing resistant BC cells. Mass spectrometry, immunofluorescence, and co-immunoprecipitation experiments elucidated ANXA6's role in enhancing PKCα/EGFR complex formation and inhibiting the EGFR pathway. ChIP-PCR and dual-luciferase assays determined PRRX1's regulatory role on ANXA6 transcription. Finally, the impact of ANXA6 in vivo was evaluated using xenograft models.

Results

Bioinformatics analyses showed a significant correlation between ANXA6 expression and cisplatin sensitivity. In vitro and in vivo experiments confirmed that ANXA6 was a new target for cisplatin treatment. ANXA6 overexpression not only enhanced cell viability inhibition, DNA damage and apoptosis caused by cisplatin, but also re-sensitized cisplatin-resistant cells. Mechanistically, ANXA6 promotes PKCα/EGFR complex formation, inhibiting EGFR phosphorylation and downstream AKT and ERK1/2. Moreover, PRRX1 was identified as a transcription factor promoting ANXA6 expression, thereby augmenting the cytotoxic effects of cisplatin.

Conclusion

Our study reveals the mechanism by which ANXA6 enhances cisplatin sensitivity and re-sensitizes resistant cells. The roles of PRRX1 and ANXA6 in cisplatin resistance offer new therapeutic targets to overcome cisplatin resistance in clinical practice.

Abstract Image

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转录因子PRRX1激活的ANXA6可促进表皮生长因子受体-PKCα复合物的形成，并增强膀胱癌对顺铂的敏感性。

背景：肿瘤对顺铂的耐药性是一项重大的临床挑战，尤其是在膀胱癌（BC）中。ANXA6是annexin家族的成员，它在顺铂耐药中的作用尚不清楚。本研究探讨了ANXA6在促进顺铂敏感性中的作用：生物信息学分析和临床标本验证评估了ANXA6与顺铂治疗之间的相关性。一系列检测方法，包括CCK-8、克隆形成检测、活性氧（ROS）和细胞凋亡流式细胞仪检测以及彗星检测，被用来证实ANXA6在增强顺铂敏感性和使耐药BC细胞再敏感中的作用。质谱分析、免疫荧光和共免疫沉淀实验阐明了ANXA6在增强PKCα/EGFR复合物形成和抑制表皮生长因子受体通路中的作用。ChIP-PCR 和双荧光素酶测定确定了 PRRX1 对 ANXA6 转录的调控作用。最后，利用异种移植模型评估了ANXA6在体内的影响：生物信息学分析表明，ANXA6的表达与顺铂敏感性之间存在显著相关性。体外和体内实验证实，ANXA6是顺铂治疗的新靶点。过表达ANXA6不仅能增强顺铂对细胞活力的抑制、DNA损伤和凋亡，还能使顺铂耐药细胞重新敏感。从机理上讲，ANXA6 可促进 PKCα/EGFR 复合物的形成，抑制表皮生长因子受体磷酸化及下游的 AKT 和 ERK1/2。此外，还发现PRRX1是促进ANXA6表达的转录因子，从而增强了顺铂的细胞毒性作用：我们的研究揭示了 ANXA6 增强顺铂敏感性并使耐药细胞再敏感的机制。PRRX1和ANXA6在顺铂耐药性中的作用为临床实践中克服顺铂耐药性提供了新的治疗靶点。

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

Life sciences 医学-药学

CiteScore

12.20

自引率

1.60%

发文量

841

审稿时长

6 months

期刊介绍： Life Sciences is an international journal publishing articles that emphasize the molecular, cellular, and functional basis of therapy. The journal emphasizes the understanding of mechanism that is relevant to all aspects of human disease and translation to patients. All articles are rigorously reviewed. The Journal favors publication of full-length papers where modern scientific technologies are used to explain molecular, cellular and physiological mechanisms. Articles that merely report observations are rarely accepted. Recommendations from the Declaration of Helsinki or NIH guidelines for care and use of laboratory animals must be adhered to. Articles should be written at a level accessible to readers who are non-specialists in the topic of the article themselves, but who are interested in the research. The Journal welcomes reviews on topics of wide interest to investigators in the life sciences. We particularly encourage submission of brief, focused reviews containing high-quality artwork and require the use of mechanistic summary diagrams.