Cao Jinlong, Chen Siyu, Wang Jirong, Fan Xinpeng, Liu Shanhui, Li Xiaoran, Yang Li
Department of Urology, The Second Hospital & Clinical Medical School, Lanzhou University, Lanzhou 730000, China; Gansu Province Clinical Research Center for Urology, Lanzhou 730000, China.
Gansu Province Clinical Research Center for Urology, Lanzhou 730000, China.
Life Sci. 2024 Dec 15;359:123228. doi: 10.1016/j.lfs.2024.123228. Epub 2024 Nov 9.
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.
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.
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.
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.
肿瘤对顺铂的耐药性是一个重大的临床挑战，尤其是在膀胱癌（BC）中。膜联蛋白A6（ANXA6）是膜联蛋白家族的成员，其在顺铂耐药中的作用尚不清楚。本研究探讨ANXA6在提高顺铂敏感性方面的作用。
通过生物信息学分析和临床标本验证评估ANXA6与顺铂治疗之间的相关性。采用一系列实验，包括CCK-8法、克隆形成实验、活性氧（ROS）和凋亡的流式细胞术检测以及彗星实验，来证实ANXA6在增强顺铂敏感性和使耐药BC细胞重新敏感方面的作用。质谱分析、免疫荧光和免疫共沉淀实验阐明了ANXA6在增强蛋白激酶Cα（PKCα）/表皮生长因子受体（EGFR）复合物形成和抑制EGFR通路中的作用。染色质免疫沉淀-聚合酶链反应（ChIP-PCR）和双荧光素酶实验确定了脯氨酸丰富的同源异型框1（PRRX1）对ANXA6转录的调控作用。最后，使用异种移植模型评估ANXA6在体内的影响。
生物信息学分析显示ANXA6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。ANXA6的过表达不仅增强了顺铂引起的细胞活力抑制、DNA损伤和凋亡，还使顺铂耐药细胞重新敏感。机制上，ANXA6促进PKCα/EGFR复合物的形成，抑制EGFR磷酸化及其下游的蛋白激酶B（AKT）和细胞外信号调节激酶1/2（ERK1/2）。此外，PRRX1被鉴定为促进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增强顺铂敏感性和使耐药细胞重新敏感的机制。PRRX1和ANXA6在顺铂耐药中的作用为临床实践中克服顺铂耐药提供了新的治疗靶点。
