下调的肿瘤相关成纤维细胞外泌体 microRNA-148b-3p 通过下调 Wnt/β-catenin 通路和上调 PTEN 增强膀胱癌细胞的化疗敏感性。

Downregulated exosomal microRNA-148b-3p in cancer associated fibroblasts enhance chemosensitivity of bladder cancer cells by downregulating the Wnt/β-catenin pathway and upregulating PTEN.

机构信息

Department of Urology, RenMin Hospital of Wuhan University, Wuhan, 430060, Hubei, People's Republic of China.

Department of Medical Laboratory Science, The Fifth People's Hospital of Wuxi, Nanjing Medical University, 1215 Guangrui Road, Jiangsu, 214000, Wuxi, People's Republic of China.

出版信息

Cell Oncol (Dordr). 2021 Feb;44(1):45-59. doi: 10.1007/s13402-020-00500-0. Epub 2021 Jan 10.

DOI:10.1007/s13402-020-00500-0

PMID:33423167

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7906940/

Abstract

OBJECTIVE

Exosomes derived from cancer-associated fibroblasts (CAFs) are known as important drivers of tumor progression. Previously, microRNA (miR)-148b-3p has been found to be upregulated in bladder cancers as well as in body fluids (blood, urine) of bladder cancer patients. Here, we aimed to explore the role of CAF-derived exosome miR-148b-3p in bladder cancer progression and chemosensitivity.

METHODS

Transwell, MTT, flow cytometry and colony formation assays were applied to assess the effects of CAF-derived exosomes on bladder cancer cell metastasis, epithelial-mesenchymal transition (EMT) and chemosensitivity. A dual luciferase reporter assay was employed to evaluate the targeting relationship between miR-148b-3p and PTEN. Gain- and loss- of function assays were conducted to explore the roles of miR-148b-3p and PTEN in the behavior of bladder cancer cells. The role of PTEN in the metastasis, EMT and chemosensitivity of bladder cancer cells was assessed both in vivo and in vitro.

RESULTS

We found that CAF-derived exosomes promoted the metastasis, EMT and drug resistance of bladder cancer cells. We also found that CAF-derived exosomes could directly transport miR-148b-3p into bladder cancer cells. In a xenograft mouse model we found that CAF-derived exosomes increased miR-148b-3p expression levels and promoted tumor proliferation, metastasis and drug resistance. PTEN was validated as a target of miR-148b-3p. Concordantly, we found that PTEN overexpression inhibited EMT, metastasis and chemoresistance in bladder cancer cells, reversing the tumor promoting effects of miR-148b-3p via the Wnt/β-catenin pathway.

CONCLUSIONS

Our results suggest that miR-148b-3p downregulation in CAF-derived exosomes, thereby inhibiting the Wnt/β-catenin pathway and promoting PTEN expression, may offer potential opportunities for bladder cancer treatment.

摘要

目的

肿瘤相关成纤维细胞（CAF）衍生的外泌体被认为是肿瘤进展的重要驱动因素。先前发现，miR-148b-3p 在膀胱癌以及膀胱癌患者的体液（血液、尿液）中上调。在这里，我们旨在探索 CAF 衍生的外泌体 miR-148b-3p 在膀胱癌进展和化疗敏感性中的作用。

方法

使用 Transwell、MTT、流式细胞术和集落形成实验来评估 CAF 衍生的外泌体对膀胱癌细胞转移、上皮-间充质转化（EMT）和化疗敏感性的影响。双荧光素酶报告实验用于评估 miR-148b-3p 与 PTEN 之间的靶向关系。进行增益和缺失功能实验以探索 miR-148b-3p 和 PTEN 在膀胱癌细胞行为中的作用。在体内和体外评估 PTEN 在膀胱癌细胞转移、EMT 和化疗敏感性中的作用。

结果

我们发现 CAF 衍生的外泌体促进了膀胱癌细胞的转移、EMT 和耐药性。我们还发现 CAF 衍生的外泌体可以直接将 miR-148b-3p 转运到膀胱癌细胞中。在异种移植小鼠模型中，我们发现 CAF 衍生的外泌体增加了 miR-148b-3p 的表达水平，并促进了肿瘤的增殖、转移和耐药性。PTEN 被验证为 miR-148b-3p 的靶标。一致地，我们发现 PTEN 过表达抑制了膀胱癌细胞的 EMT、转移和化疗耐药性，通过 Wnt/β-catenin 通路逆转了 miR-148b-3p 的肿瘤促进作用。