HNF1A-AS1/miR-92a-3p轴通过竞争性调节JNK通路影响非小细胞肺癌的放射敏感性。
The HNF1A-AS1/miR-92a-3p axis affects the radiosensitivity of non-small cell lung cancer by competitively regulating the JNK pathway.
作者信息
Wang Zhiyu, Liu Liang, Du Yuankun, Mi Yuan, Wang Lei
机构信息
Department of Oncology immunology, Fourth Hospital of Hebei Medical University, Shijiazhuang City, Hebei Province, 050011, People's Republic of China.
Tumor Institute, The Fourth Hospital of Hebei Medical University, Shijiazhuang City, Hebei Province, 050011, People's Republic of China.
出版信息
Cell Biol Toxicol. 2021 Oct;37(5):715-729. doi: 10.1007/s10565-021-09595-z. Epub 2021 Mar 23.
BACKGROUND
It has been widely reported that long non-coding RNAs (lncRNAs) could affect the varieties of tumor response to radiotherapy. LncRNA HNF1A-AS1 is transcribed from HNF1A gene cluster's antisense strand. This work focused on the mechanism of how HNF1A-AS1 participated in the radiosensitivity of non-small cell lung cancer (NSCLC).
METHODS
The mRNA or protein expression of HNF1A-AS1, miR-92a-3p MAP2K4, and JNK in NSCLC cells and tissues was detected by qRT-PCR or western blotting. RNA immunoprecipitation (RIP) detection and luciferase reporting system were used to evaluate the relationship between HNFA-AS1 and miR-92a-3p or between miR-92a-3p and MAP2K4. Flow cytometry assays, colony formation, and MTT were performed to analyze the function changes in A549 and Calu-1 cells. The rescue experiment was also conducted to explore the underlying mechanisms.
RESULTS
HNF1A-AS1 was investigated in NSCLC cells and tissues and highly related to the advanced pathological stage. HNF1A-AS1 bound with miR-92a-3p, which was downregulated in NSCLC. It showed that miR-92a-3p was negatively related to HNF1A-AS1. Knockdown of HNF1A-AS1 impacted most cell biological behaviors in NSCLC cells, including restricting the proliferation and aggravating apoptosis. Furthermore, knockdown of HNF1A-AS1 dramatically enhanced radiotherapy sensitivity of NSCLC. Moreover, miR-92a-3p was found to target MAP2K4 and could reduce MAP2K4 expression. Inhibition of HNF1A-AS1 elevated radiotherapy sensitivity and retarded the progression of NSCLC cells, followed by decreasing expression levels of MAP2K4. Besides, MAP2K4 mimic rescued the si-HNF1A-AS1 effects on the biological behavior of NSCLC cells.
CONCLUSION
HNF1A-AS1 is highly expressed in NSCLC. MiR-92a-3p is the target gene of HNF1A-AS1 and involved in tumor progression by regulating the MAP2K4/JNK pathway. HNF1AS1/miR-92a-3p/MAP2K4 axis plays important roles in radiotherapy resistance of NSCLC.
背景
长期以来，广泛报道称长链非编码RNA（lncRNA）可影响肿瘤对放疗反应的多种情况。LncRNA HNF1A-AS1从HNF1A基因簇的反义链转录而来。本研究聚焦于HNF1A-AS1参与非小细胞肺癌（NSCLC）放射敏感性的机制。
方法
采用qRT-PCR或蛋白质免疫印迹法检测NSCLC细胞和组织中HNF1A-AS1、miR-92a-3p、MAP2K4和JNK的mRNA或蛋白表达。运用RNA免疫沉淀（RIP）检测和荧光素酶报告系统评估HNFA-AS1与miR-92a-3p之间或miR-92a-3p与MAP2K4之间的关系。通过流式细胞术检测、集落形成实验和MTT实验分析A549和Calu-1细胞的功能变化。还进行了挽救实验以探究潜在机制。
结果
在NSCLC细胞和组织中对HNF1A-AS1进行了研究，发现其与晚期病理阶段高度相关。HNF1A-AS1与miR-92a-3p结合，miR-92a-3p在NSCLC中表达下调。结果表明miR-92a-3p与HNF1A-AS1呈负相关。敲低HNF1A-AS1影响NSCLC细胞的大多数细胞生物学行为，包括抑制增殖和加剧凋亡。此外，敲低HNF1A-AS1显著增强了NSCLC的放射敏感性。而且，发现miR-92a-3p靶向MAP2K4并可降低MAP2K4表达。抑制HNF1A-AS1可提高放射敏感性并延缓NSCLC细胞的进展，随后降低MAP2K4的表达水平。此外，MAP2K4模拟物挽救了si-HNF1A-AS1对NSCLC细胞生物学行为的影响。
结论
HNF1A-AS1在NSCLC中高表达。miR-92a-3p是HNF1A-AS1的靶基因，通过调节MAP2K4/JNK途径参与肿瘤进展。HNF1AS1/miR-92a-3p/MAP2K4轴在NSCLC的放疗抵抗中起重要作用。
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