Chen Wenyu, Tan Xiaoli, Yang Qi, Fang Zhixian, Xu Yufen
Department of Respiratory, Affiliated Hospital of Jiaxing University, Jiaxing, Zhejiang 314000, China.
Department of Respiratory, Affiliated Hospital of Jiaxing University, Jiaxing, Zhejiang 314000, China.
Cell Signal. 2022 Jun;94:110326. doi: 10.1016/j.cellsig.2022.110326. Epub 2022 Mar 31.
MALAT1 has been implicated in tumor progression. But the mechanism and role underlying MALAT1 in non-small cell lung cancer (NSCLC) cell resistance to gemcitabine (GEM) remain rarely understood.
Through bioinformatics analysis, we predicted MALAT1/miR-27a-5p/PBOV1 regulatory axis and constructed GEM resistant A549/GEM cell line, and A549 was the parent cell line. qRT-PCR was utilized to assess MALAT1, miR-27a-5p and PBOV1 expression in A549 and A549/GEM cells. MTT method and colony formation assay were utilized to measure cell viability and cell proliferation. Flow cytometry was conducted to assess cell cycle and cell apoptosis. Wound healing and Transwell assays were conducted to measure cell migratory and invasive potentials. Dual-luciferase reporter gene assay and RNA immunoprecipitation were utilized to identify the targeted relationship between MALAT1 and miR-27a-5p, and the former assay was also utilized to determine the targeted relationship between miR-27a-5p and PBOV1. The impacts of MALAT1/miR-27a-5p/PBOV1 on tumor growth and GEM resistance of NSCLC cells in vivo were validated by using the tumor xenograft model.
MALAT1 was observed to be highly expressed in tissues and cells of GEM resistant patients. Forced level of MALAT1 could markedly enhance A549 cell resistance to GEM, but this impact could be weakened by silencing MALAT1. MALAT1 downregulated miR-27a-5p level. PBOV1 was the target of miR-27a-5p and could significantly enhance GEM resistance of NSCLC cell. MALAT1 facilitated tumor growth in vivo via targeting miR-27a-5p/PBOV1 and enhanced resistance of NSCLC cells to GEM.
MALAT1/miR-27a-5p/PBOV1 axis was implicated in NSCLC cell resistance to GEM. We deepened our understanding about how MALAT1 enhanced NSCLC cell resistance to GEM and provided development of therapeutic strategy for NSCLC with a possible target.
MALAT1与肿瘤进展有关。但MALAT1在非小细胞肺癌（NSCLC）细胞对吉西他滨（GEM）耐药中的机制和作用仍鲜为人知。
通过生物信息学分析，我们预测了MALAT1/miR-27a-5p/PBOV1调控轴，并构建了GEM耐药A549/GEM细胞系，A549为亲本细胞系。采用qRT-PCR评估A549和A549/GEM细胞中MALAT1、miR-27a-5p和PBOV1的表达。采用MTT法和集落形成试验检测细胞活力和细胞增殖。通过流式细胞术评估细胞周期和细胞凋亡。采用伤口愈合试验和Transwell试验检测细胞迁移和侵袭能力。采用双荧光素酶报告基因试验和RNA免疫沉淀法鉴定MALAT1与miR-27a-5p之间的靶向关系，前者还用于确定miR-27a-5p与PBOV1之间的靶向关系。通过肿瘤异种移植模型验证MALAT1/miR-27a-5p/PBOV1对NSCLC细胞肿瘤生长和GEM耐药的影响。
观察到MALAT1在GEM耐药患者的组织和细胞中高表达。强制提高MALAT1水平可显著增强A549细胞对GEM的耐药性，但沉默MALAT1可减弱这种影响。MALAT1下调miR-27a-5p水平。PBOV1是miR-27a-5p的靶标，可显著增强NSCLC细胞对GEM的耐药性。MALAT1通过靶向miR-27a-5p/PBOV1促进体内肿瘤生长，并增强NSCLC细胞对GEM的耐药性。
MALAT1/miR-27a-5p/PBOV1轴与NSCLC细胞对GEM的耐药有关。我们加深了对MALAT1如何增强NSCLC细胞对GEM耐药性的理解，并为NSCLC的治疗策略开发提供了一个可能的靶点。
