MYH11对NUPR1的转录调控激活PI3K/AKT并通过铁死亡和巨噬细胞M2极化促进膀胱癌进展。

Transcriptional Regulation of NUPR1 by MYH11 Activates PI3 K/AKT and Promotes Bladder Cancer Progression Through Ferroptosis and M2 Polarization of Macrophages.

作者信息

Zhang Lifeng, Zhang Li, Shi Zebin, Mi Yuanyuan, Zhang Lei, Shi Xiaokai, Gao Shenglin, Zuo Li

机构信息

Department of Urology, The Affiliated Changzhou Second People's Hospital of Nanjing Medical University (The Third Affiliated Hospital of Nanjing Medical University), Changzhou, China.

Department of Urology, Changzhou Second People's Hospital, Changzhou Medical Center, Nanjing Medical University, Changzhou, China.

出版信息

Technol Cancer Res Treat. 2025 Jan-Dec;24:15330338241305434. doi: 10.1177/15330338241305434.

DOI:10.1177/15330338241305434

PMID:39962891

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11833819/

Abstract

BACKGROUND

NUPR1 is a small molecule protein that plays an important role in tumor progression and drug resistance. Our previous study found that NUPR1 promotes the progression of bladder cancer, but the specific mechanism is still unclear. MYH11 encodes the smooth muscle myosin heavy chain and belongs to the conventional myosin family. MYH11 has been found to be associated with a variety of malignant tumors.

METHODS

We identified MYH11 as an upstream regulator of NUPR1 using a bioinformatics approach and tested this hypothesis by knocking down MYH11 and ChIP-qPCR. Subsequently, we verified the association of MYH11 and NUPR1 with the PI3 K/AKT pathway by WB. In addition, gene enrichment results showed that the effect of NUPR1 on bladder cancer was related to ferroptosis and M2 macrophage polarization. We examined ferroptosis metabolites in bladder cancer cells overexpressing NUPR1 and expression of the M2 macrophage marker CD206 in NUPR1 overexpression or MYH11 knockdown bladder cancer cells.

RESULTS

Bioinformatics results showed that MYH11 was positively correlated with NUPR1, and there may be a mutual binding site at the promoter of NUPR1. Knockdown of MYH11 decreased NUPR1 expression, and ChIP-qPCR showed that MYH11 bound to the promoter of NUPR1. Subsequently, WB results showed that MYH11 knockdown inhibited the PI3 K/AKT pathway, whereas NUPR1 overexpression activated this pathway. After adding ferroptosis activator, the viability of bladder cancer cells decreased, and the content of Fe and MDA increased. However, ferroptosis was significantly inhibited after overexpression of NUPR1. Knockdown of MYH11 inhibited M2 macrophage polarization, while overexpression of NUPR1 promoted this process.

CONCLUSION

This study suggests that MYH11 activates the PI3 K/AKT pathway by up-regulating the expression of NUPR1, and promotes bladder cancer progression by inhibiting ferroptosis and promoting M2 polarization of macrophages.

摘要

背景

NUPR1是一种小分子蛋白，在肿瘤进展和耐药性中起重要作用。我们之前的研究发现NUPR1促进膀胱癌进展，但具体机制仍不清楚。MYH11编码平滑肌肌球蛋白重链，属于传统肌球蛋白家族。已发现MYH11与多种恶性肿瘤有关。

方法

我们使用生物信息学方法将MYH11鉴定为NUPR1的上游调节因子，并通过敲低MYH11和ChIP-qPCR验证这一假设。随后，我们通过蛋白质免疫印迹法（WB）验证MYH11和NUPR1与PI3K/AKT通路的关联。此外，基因富集结果表明NUPR1对膀胱癌的影响与铁死亡和M2巨噬细胞极化有关。我们检测了过表达NUPR1的膀胱癌细胞中的铁死亡代谢物，以及NUPR1过表达或MYH11敲低的膀胱癌细胞中M2巨噬细胞标志物CD206的表达。

结果

生物信息学结果显示MYH11与NUPR1呈正相关，且在NUPR1启动子处可能存在相互结合位点。敲低MYH11可降低NUPR1表达，ChIP-qPCR显示MYH11与NUPR1启动子结合。随后，WB结果显示敲低MYH11可抑制PI3K/AKT通路，而过表达NUPR1可激活该通路。添加铁死亡激活剂后，膀胱癌细胞活力下降，铁和丙二醛（MDA）含量增加。然而，NUPR1过表达后铁死亡明显受到抑制。敲低MYH11可抑制M2巨噬细胞极化，而过表达NUPR1则促进这一过程。