Exosomal transfer of p-STAT3 promotes acquired 5-FU resistance in colorectal cancer cells
Exosomal transfer of p-STAT3 promotes acquired 5-FU resistance in colorectal cancer cells
复制标题
p-STAT3 的外泌体转移促进结直肠癌细胞获得性 5-FU 耐药性
DOI：
10.1186/s13046-019-1314-9
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发表时间：
2019
影响因子：
11.3
通讯作者：
Liu Huanliang
中科院分区：
文献类型：
--
作者：
Zhang Qian;Liu Rui Xian;Chan Ka Wo;Hu Jiancong;Zhang Jingdan;Wei Lili;Tan Huiliu;Yang Xiangling;Liu Huanliang
获得性耐药仍然限制了5-氟尿嘧啶(5-FU)的临床应用。由于Exosome是参与细胞间通讯的重要囊泡，因此它们在获得性5-FU耐药中的作用还有待阐明。本研究旨在探讨5-FU耐药细胞外切体(RKO/R)维持敏感细胞获得性5-FU耐药(RKO/P)的机制。采用实时细胞分析(RTCA)和流式细胞仪检测细胞存活率和细胞凋亡率。使用鸟枪式蛋白质组学进行外体蛋白图谱分析。应用抑制剂和siRNAs研究所选蛋白对5-FU耐药的影响。通过Western blotting和高含量分析检测外体p-STAT3(Tyr705)对caspase级联的影响。建立异种移植模型，观察外切体p-STAT3能否在体内诱导5-FU耐药。结果RKO/R细胞外切体在5-FU处理过程中显著促进细胞存活。蛋白质组学和WB分析结果表明，GSTP1和p-STAT3(Tyr705)富含在RKO/R细胞的外切体中。P-STAT3再敏化RKO/P细胞对5-FU的抑制作用RKO/R细胞外切体包装的p-STAT3可提高肿瘤细胞对5-FU的体内耐药性。结论含p-STAT3的外切体参与了结直肠癌获得性5-FU耐药的新机制。这项研究为增强5-FU的反应和寻找化疗耐药的生物标志物提供了一种新的选择。
BackgroundAcquired resistance remains a limitation of the clinical use of 5-fluorouracil (5-FU). Because exosomes, are important vesicles participating in intercellular communication, their contribution to the development of acquired 5-FU resistance needs to be elucidated. In this study, we aimed to examine the underlying mechanisms of exosomes from 5-FU resistant cells (RKO/R) in sustaining acquired 5-FU resistance in sensitive cells (RKO/P).MethodsExosomes from a 5-FU-resistant cell line (RKO/R) and its parental cell line RKO/P were isolated and co-cultured with 5-FU-sensitive cells. Real-time cellular analysis (RTCA) and FACS analysis were used to examine cell viability and apoptosis. Exosomal protein profiling was performed using shotgun proteomics. Inhibitors and siRNAs were applied to study the involvement of selected proteins in 5-FU resistance. The effect of exosomal p-STAT3 (Tyr705) on the caspase cascade was examined by western blotting (WB) and high content analysis. Xenograft models were established to determine whether exosomal p-STAT3 can induce 5-FU resistance in vivo.ResultsOur results indicated that exosomes from RKO/R cells significantly promoted cell survival during 5-FU treatment. Proteomics and WB analysis results indicated that GSTP1 and p-STAT3 (Tyr705) were enriched in exosomes from RKO/R cells. Inhibition of p-STAT3 re-sensitized RKO/P cells to 5-FU via caspase cascade. Furthermore, p-STAT3 packaged by exosomes from RKO/R cells increased resistance of tumor cells to 5-FU in vivo.ConclusionsOur results reveal a novel mechanism by which p-STAT3-containing exosomes contribute to acquired 5-FU resistance in CRC. This study suggests a new option for potentiating the 5-FU response and finding biomarkers for chemotherapy resistance.