Non-glycanated ΔDCN isoform in muscle invasive bladder cancer mediates cancer stemness and gemcitabine resistance.

IF 4.9 2区医学 Q2 CELL BIOLOGY Cellular Oncology Pub Date : 2024-10-28 DOI:10.1007/s13402-024-00998-8

Nisha Wu, Jinxiang Wang, Mingming Fan, Yanling Liang, Xiao Wei Qi, Fan Deng, Fangyin Zeng

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Abstract

Background: The small leucine-rich proteoglycan decorin (DCN) is recognized for its diverse roles in tissue homeostasis and malignant progression. Nevertheless, the regulatory effects of DCN on bladder cancer stem cells (BCSCs) and the underlying mechanisms in muscle-invasive bladder cancer (MIBC) remain to be elucidated.

Methods: The study obtained data (including scRNA-seq, clinicopathological characteristics, and survival) were acquired from TCGA and GEO. The BCSCs were cultured by enriching the suspension culture in a serum-free medium, followed by flow cytometry sorting. Overexpression/knockdown was constructed by utilizing lentivirus. The surface biomarkers of cancer stem cells were identified via flow cytometry. Cell proliferation and self-renewal were evaluated by CCK8 and Sphere formation assays, and in vivo tumor growth was evaluated with subcutaneous xenografts.

Results: Total DCN expression was significantly elevated in muscle-invasive bladder cancer (MIBC) and was associated with poor prognosis. The ΔDCN isoform, which lacks glycosylation sites, was identified in bladder cancer stem cells (BCSCs) derived from clinical tissue samples and bladder cancer cell lines. Suppression of ΔDCN expression resulted in a reduction of BCSC stemness. Both in vitro and in vivo experiments indicated that overexpression of full-length DCN inhibited stemness within the extracellular matrix. Conversely, overexpression of ΔDCN and the introduction of exogenous recombinant decorin protein in ΔDCN-knockdown BCSC-SW780 cell lines enhanced stemness within the cytoplasm. The ΔDCN isoform exhibited resistance to gemcitabine chemotherapy in vitro.

Conclusion: Non-glycanated ΔDCN isoforms were identified in bladder cancer stem cells (BCSCs), where they exhibited differential cytoplasmic localization and promoted oncogenic effects by inducing a stemness phenotype and conferring resistance to gemcitabine chemotherapy. These oncogenic effects are in stark contrast to the anti-tumor functions of glycosylated DCN in the extracellular matrix. The ratio of ΔDCN isoforms to glycosylated DCN is pivotal in predicting tumor progression and therapeutic resistance.

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肌浸润性膀胱癌中的非糖化ΔDCN同工酶介导癌症干性和吉西他滨耐药性。

背景：富含亮氨酸的小蛋白多糖decorin（DCN）被认为在组织稳态和恶性进展中发挥着多种作用。然而，DCN对膀胱癌干细胞（BSCs）的调控作用及其在肌肉浸润性膀胱癌（MIBC）中的潜在机制仍有待阐明：研究获得的数据（包括 scRNA-seq、临床病理特征和存活率）来自 TCGA 和 GEO。在无血清培养基中富集悬浮培养的 BCSCs，然后进行流式细胞术分选。利用慢病毒构建过表达/基因敲除模型。通过流式细胞术鉴定癌症干细胞的表面生物标志物。细胞增殖和自我更新通过CCK8和球形成试验进行评估，体内肿瘤生长通过皮下异种移植进行评估：结果：在肌浸润性膀胱癌（MIBC）中，DCN总表达量明显升高，且与预后不良有关。在来自临床组织样本和膀胱癌细胞系的膀胱癌干细胞（BCSCs）中发现了缺乏糖基化位点的ΔDCN异构体。抑制ΔDCN的表达可降低膀胱癌干细胞的干性。体外和体内实验都表明，过表达全长DCN会抑制细胞外基质中的干细胞。相反，在ΔDCN敲除的BCSC-SW780细胞系中，过表达ΔDCN和引入外源重组decorin蛋白可增强细胞质内的干性。ΔDCN异构体在体外表现出对吉西他滨化疗的耐药性：结论：在膀胱癌干细胞（BCSCs）中发现了非糖化ΔDCN异构体，它们表现出不同的细胞质定位，并通过诱导干性表型和赋予吉西他滨化疗抗性来促进致癌效应。这些致癌作用与细胞外基质中糖基化 DCN 的抗肿瘤功能形成了鲜明对比。ΔDCN异构体与糖基化DCN的比例在预测肿瘤进展和治疗耐药性方面至关重要。

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

Cellular Oncology ONCOLOGY-CELL BIOLOGY

CiteScore

10.30

自引率

1.50%

发文量

审稿时长

12 months

期刊介绍： The Official Journal of the International Society for Cellular Oncology Focuses on translational research Addresses the conversion of cell biology to clinical applications Cellular Oncology publishes scientific contributions from various biomedical and clinical disciplines involved in basic and translational cancer research on the cell and tissue level, technical and bioinformatics developments in this area, and clinical applications. This includes a variety of fields like genome technology, micro-arrays and other high-throughput techniques, genomic instability, SNP, DNA methylation, signaling pathways, DNA organization, (sub)microscopic imaging, proteomics, bioinformatics, functional effects of genomics, drug design and development, molecular diagnostics and targeted cancer therapies, genotype-phenotype interactions. A major goal is to translate the latest developments in these fields from the research laboratory into routine patient management. To this end Cellular Oncology forms a platform of scientific information exchange between molecular biologists and geneticists, technical developers, pathologists, (medical) oncologists and other clinicians involved in the management of cancer patients. In vitro studies are preferentially supported by validations in tumor tissue with clinicopathological associations.