lncRNA FOXD2-AS1 confers cisplatin resistance of non-small-cell lung cancer via regulation of miR 185-5p-SIX1 axis
lncRNA FOXD2-AS1 confers cisplatin resistance of non-small-cell lung cancer via regulation of miR 185-5p-SIX1 axis
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DOI：
10.2147/ott.s197454
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发表时间：
2019-01-01
影响因子：
4
通讯作者：
Li, Han-Jie
中科院分区：
文献类型：
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作者：
Ge, Peng;Cao, Lei;Li, Han-Jie
背景：化疗耐药是非小细胞肺癌（NSCLC）化疗失败的主要障碍。lncRNA是多种癌症中的一类关键调节因子，并且lncRNA FOXD 2-AS 1与NSCLC的进展有关。方法：采用逆转录qPCR方法检测FOXD 2-AS 1、miR 185 - 5 p和SIX 1 mRNA的表达水平。CCK-8法检测A549/DDP和H1299/DDP耐药细胞的增殖和化疗敏感性。采用菌落形成试验检测菌落数。transwell法检测细胞迁移和侵袭能力。Western blot检测LRP、Pgp、MRP 1和SIX 1蛋白表达水平。通过生物信息学、双荧光素酶和RNA免疫沉淀试验验证FOXD 2-AS 1与miR 185 - 5 p或miR 185 - 5 p与SIX 1之间的相关性。结果：DDP耐药NSCLC组织中FOXD 2-AS 1和SIX 1表达上调，miR 185 - 5 p表达下调。FOXD 2-AS 1基因缺失可增强A549/DDP和H1299/DDP细胞的药物敏感性，表现为集落形成、细胞增殖、迁移、侵袭和耐药相关蛋白表达减少。FOXD 2-AS 1作为miR 185 - 5 p的分子海绵，解除miR 185 - 5 p与其靶基因SIX 1的结合，导致SIX 1的去阻遏。在A549/DDP和H1299/DDP细胞中。拯救实验验证了FOXD 2-AS 1、miR 185 - 5 p和SIX 1之间的功能性相互作用。结论：FOXD 2-AS 1/miR 185 - 5 p/SIX 1可调控顺铂耐药NSCLC的生长和化疗敏感性，有望成为顺铂耐药NSCLC的治疗靶点。
Background: Chemoresistance is a major obstacle for chemotherapy failure in non-small-cell lung cancer (NSCLC). lncRNAs are a class of pivotal regulators in various cancers, and the lncRNA FOXD2-AS1 is implicated in the progression of NSCLC. However, it is still unclear whether it regulates chemosensitivity.Methods: Expression levels of FOXD2-AS1, miR185-5p, and SIX1 mRNA were identified by reverse-transcription qPCR. CCK8 assay was performed to assess cell proliferation and chemosensitivity of cisplatin-resistant A549/DDP and H1299/DDP cells. Colony-forming assay was utilized to detect colony numbers. Cell migration and invasion ability were measured by transwell assay. The protein levels of LRP, Pgp, MRP1, and SIX1 were examined by Western blot assay. The correlation between FOXD2-AS1 and miR185-5p or miR185-5p and SIX1 were validated by bioinformatic, dual-luciferase, and RNA immunoprecipitation assays. Tumor xenografts were constructed to confirm the function and mechanism of FOXD2-AS1 in chemosensitivity of DDP-resistant NSCLC.Results: FOXD2-AS1 and SIX1 were upregulated and miR185-5p downregulated in DDP-resistant NSCLC. Absence of FOXD2-AS1 enhanced drug sensitivity of A549/DDP and H1299/DDP cells, reflected by the reduced colony formation, cell proliferation, migration, invasion, and drug resistance-associated protein expression. FOXD2-AS1 acted as a molecular sponge for miR185-5p and relieved the binding of miR185-5p and its target gene SIX1, leading to the derepression of SIX1. in A549/DDP and H1299/DDP cells. Rescue experiments validated the functional interaction among FOXD2-AS1, miR185-5p, and SIX1. Moreover, FOXD2-AS1 interference receded the growth of DDP-resistant NSCLC tumors in vivo.Conclusion: FOXD2-AS1/miR185-5p/SIX1 regulates the progression and chemosensitivity of DDP-resistant NSCLC, suggesting a potential therapeutic target for cisplatin-resistant NSCLC patients.