Exosomal Transfer Of Cisplatin-Induced miR-425-3p Confers Cisplatin Resistance In NSCLC Through Activating Autophagy
Exosomal Transfer Of Cisplatin-Induced miR-425-3p Confers Cisplatin Resistance In NSCLC Through Activating Autophagy
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顺铂诱导的 miR-425-3p 的外泌体转移通过激活自噬赋予 NSCLC 顺铂耐药性
DOI：
10.2147/ijn.s221383
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发表时间：
2019-01-01
影响因子：
8
通讯作者：
Shen, Yan
中科院分区：
文献类型：
--
作者：
Ma, Yuzhu;Yuwen, Daolu;Shen, Yan
简介 外泌体是细胞间通讯的重要介质。此前，我们将循环外泌体 miR-425-3p 作为一种非侵入性预后标志物，用于预测非小细胞肺癌（NSCLC）患者对铂类化疗的临床反应。方法 通过 qRT-PCR 在 NSCLC 患者铂类化疗过程中的配对血清样本中验证循环外泌体 miR-425-3p。进行细胞共培养以检查外泌体 miR-425-3p 对受体 A549 细胞对顺铂敏感性的影响。使用生物信息学、ChIP 和荧光素酶报告基因检测，鉴定了 miR-425-3p 表达所必需的转录因子。通过蛋白质印迹和荧光显微镜测定受体细胞中的自噬活性。结果 与基线患者相比，耐受患者的血清样本中发现外泌体 miR-425-3p 水平更高。化疗期间循环外泌体 miR-425-3p 的表达呈上升趋势。此外，顺铂可以诱导 NSCLC 细胞中外泌体 miR-425-3p 的表达。从顺铂治疗或顺铂耐药的 NSCLC 细胞中分离的外泌体以 miR-425-3p 依赖性方式赋予敏感 A549 细胞化疗耐药性。发现顺铂诱导的 c-Myc 直接结合 miR-425-3p 启动子并反式激活其表达。外泌体 miR-425-3p 通过靶向 AKT1 促进受体细胞的自噬激活，最终导致化疗耐药。讨论 我们的结果表明，除了治疗反应的预后标志物之外，外泌体 miR-425-3p 可能是一种潜在的动态生物标志物，可在治疗期间调整 NSCLC 患者的顺铂耐药性，并代表治疗耐药 NSCLC 的有希望的治疗靶点。
Introduction Exosomes are important mediators of intercellular communication. Previously, we characterized circulating exosomal miR-425-3p as a non-invasive prognostic marker for predicting clinical response to platinum-based chemotherapy in patients with non-small cell lung cancer (NSCLC). Methods Circulating exosomal miR-425-3p was validated by qRT-PCR in paired serum samples from NSCLC patients during the course of platinum-based chemotherapy. Cell coculture was performed to examine the effects of exosomal miR-425-3p on the sensitivity of recipient A549 cells to cisplatin. Using bioinformatics, ChIP and luciferase reporter assays, the transcription factor essential for miR-425-3p expression was identified. Autophagic activity in the recipient cells was determined by Western blot and fluorescence microscopy. Results Higher levels of exosomal miR-425-3p were found in serum samples from the patients in tolerance versus those at baseline. An upward trend in the expression of circulating exosomal miR-425-3p was revealed during chemotherapy. Furthermore, the expression of exosomal miR-425-3p could be induced by cisplatin in NSCLC cells. Exosomes isolated from either cisplatin-treated or cisplatin-resistant NSCLC cells conferred chemoresistance to sensitive A549 cells in a miR-425-3p-dependent manner. Cisplatin-induced c-Myc was found to directly bind the miR-425-3p promoter and transactivated its expression. Exosomal miR-425-3p facilitated autophagic activation in the recipient cells by targeting AKT1, eventually leading to chemoresistance. Discussion Our results suggest that apart from a prognostic marker of treatment response, exosomal miR-425-3p might be a potential dynamic biomarker to tailor cisplatin resistance in NSCLC patients during the treatment and represent a promising therapeutic target for therapy-resistant NSCLC.