The PVT1/miR-216b/Beclin-1 regulates cisplatin sensitivity of NSCLC cells via modulating autophagy and apoptosis
The PVT1/miR-216b/Beclin-1 regulates cisplatin sensitivity of NSCLC cells via modulating autophagy and apoptosis
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DOI：
10.1007/s00280-019-03808-3
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发表时间：
2019-05-01
影响因子：
3
通讯作者：
Chen, Liangxin
中科院分区：
文献类型：
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作者：
Chen, Liangfeng;Han, Xiaobing;Chen, Liangxin
目的非小细胞肺癌（NSCLC）化疗耐药的原因是以顺铂为基础的化疗方案的疗效仍是一个悬而未决的问题。本研究旨在探讨长链非编码RNA浆细胞瘤变异易位1（PVT 1）在非小细胞肺癌顺铂敏感性中的作用及其机制。根据患者的临床病理参数，探讨PVT 1的临床价值。顺铂敏感或耐药细胞（A549或A549/DDP）用于体外实验。通过细胞活力、凋亡、自噬和动物实验研究顺铂敏感性。分别采用定量逆转录聚合酶链反应（qRT-PCR）和western blot检测PVT 1、microRNA-216 b（miR-216 b）和凋亡或自噬相关蛋白的表达。采用荧光素酶报告基因分析和RNA免疫沉淀（RIP）技术检测miR-216 b与PVT 1和Beclin-1的相互作用。结果PVT 1在NSCLC中高表达，且与NSCLC患者的不良预后相关（P <0. 05）。在A549/DDP细胞中，PVT 1基因敲减可增强顺铂诱导的细胞活力抑制和凋亡诱导作用，而加入PVT 1则相反（*P < 0.05，P-# < 0.05）。此外，PVT 1的积累促进了NSCLC细胞的自噬和体内肿瘤生长（*P < 0.05，P-# < 0.05）。此外，miR-216 b与PVT 1或Beclin-1相互作用。Beclin-1逆转了miR-216 b介导的对NSCLC细胞自噬和凋亡的作用（*P < 0.05，P-# < 0.05）。结论PVT 1可能通过miR-216 b/Beclin-1途径调控细胞凋亡和自噬，从而与miR-216 b竞争性抑制NSCLC顺铂敏感性，为提高NSCLC化疗疗效提供了新的靶点。
Purpose The efficacy of cisplatin-based chemotherapy remains an open question for chemo-resistance in non-small cell lung cancer (NSCLC). This study aimed to explore the role and mechanism of long noncoding RNA plasmacytoma variant translocation 1 (PVT1) in cisplatin sensitivity of NSCLC.Methods Paired tumor and adjacent tissues were collected from forty patients with NSCLC. The clinical value of PVT1 was investigated according to clinicopathological parameters of patients. Cisplatin-sensitive or -resistant cells (A549 or A549/DDP) were used for in vitro experiments. Cell viability, apoptosis, autophagy and animal experiments were conducted to investigate cisplatin sensitivity. The expressions of PVT1, microRNA-216b (miR-216b) and apoptosis- or autophagy-related proteins were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) or western blot assay, respectively. Luciferase reporter assay and RNA immunoprecipitation (RIP) assay were conducted to probe the interaction between miR-216b and PVT1 or Beclin-1.Results PVT1 was highly expressed and associated with poor prognosis of NSCLC patients (*P < 0.05). PVT1 knockdown enhanced cisplatin-induced viability inhibition and apoptosis induction in A549/DDP cells, but addition of PVT1 caused an opposite effect in A549 cells (*P < 0.05, P-# < 0.05). Moreover, accumulation of PVT1 facilitated autophagy of NSCLC cells and tumor growth in vivo (*P < 0.05, P-# < 0.05). In addition, miR-216b interacted with PVT1 or Beclin-1. Beclin-1 reversed miR-216b-mediated effect on autophagy and apoptosis of NSCLC cells (*P < 0.05, P-# < 0.05). Besides, Beclin-1 protein expression was regulated by PVT1 and miR-216b (*P < 0.05, P-# < 0.05).Conclusions PVT1 may function as a competing endogenous RNA for miR-216b to inhibit cisplatin sensitivity of NSCLC through regulating apoptosis and autophagy via miR-216b/Beclin-1 pathway, providing a novel target for improving chemotherapy efficacy of NSCLC.