MCF2L-AS1/miR-874-3p/STAT3反馈环促进肺腺癌细胞生长和顺铂耐药。

MCF2L-AS1/miR-874-3p/STAT3 feedback loop contributes to lung adenocarcinoma cell growth and cisplatin resistance.

作者信息

Xu Min, Zheng Jing, Wang Jun, Huang Haitao, Hu Gang, He Hailan

机构信息

Department of Respiratory and Critical Care Medicine, the Fifth People's Hospital of Chengdu, China.

Chengdu Women's and Children's Central Hospital, China.

出版信息

Heliyon. 2023 Oct 29;9(11):e21342. doi: 10.1016/j.heliyon.2023.e21342. eCollection 2023 Nov.

DOI:10.1016/j.heliyon.2023.e21342

PMID:37954302

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10637966/

Abstract

BACKGROUND

Long noncoding RNA (lncRNA) is widely acknowledged for its crucial role in the biological processes of various human cancers. MCF2L antisense RNA 1 (MCF2L-AS1) is a newly identified lncRNA, which remains unexplored in the context of cancer.

METHODS

MCF2L-AS1 expression was examined using qRT-PCR analysis. The impact of MCF2L-AS1 on LUAD cell growth was assessed through CCK-8, colony formation, EdU, caspase-3 activity, TUNEL, Western blot, and transwell assays. The interaction between miR-874-3p and MCF2L-AS1 or STAT3 was confirmed by RIP, luciferase reporter, and RNA pull-down assays.

RESULTS

Our study demonstrated the overexpression of MCF2L-AS1 in LUAD cells. Functionally, the silencing of MCF2L-AS1 hindered cell proliferation, migration, and invasion, while promoting cell apoptosis. Notably, the depletion of MCF2L-AS1 was associated with decreased cisplatin resistance. Mechanistically, MCF2L-AS1 was identified as an upstream gene of miR-874-3p, negatively regulating its expression. Following this, STAT3, the downstream target of miR-874-3p, was identified. Additionally, the expression of STAT3 was inversely related to miR-874-3p and positively regulated by MCF2L-AS1. A restoration assay suggested that MCF2L-AS1 promoted LUAD cell growth by sponging miR-874-3p and modulating STAT3 expression. Intriguingly, STAT3 was subsequently confirmed as a transcription factor that binds to the MCF2L-AS1 promoter, thereby enhancing its transcription.

CONCLUSIONS

The MCF2L-AS1/miR-874-3p/STAT3 feedback loop plays a significant role in LUAD cell growth and cisplatin resistance.

摘要

背景

长链非编码RNA（lncRNA）因其在多种人类癌症生物学过程中的关键作用而被广泛认可。MCF2L反义RNA 1（MCF2L-AS1）是一种新发现的lncRNA，在癌症背景下尚未得到研究。

方法

采用qRT-PCR分析检测MCF2L-AS1的表达。通过CCK-8、集落形成、EdU、caspase-3活性、TUNEL、蛋白质免疫印迹和transwell实验评估MCF2L-AS1对肺腺癌（LUAD）细胞生长的影响。通过RNA免疫沉淀（RIP）、荧光素酶报告基因和RNA下拉实验证实miR-874-3p与MCF2L-AS1或信号转导和转录激活因子3（STAT3）之间的相互作用。

结果

我们的研究证明MCF2L-AS1在LUAD细胞中过表达。在功能上，MCF2L-AS1的沉默阻碍了细胞增殖、迁移和侵袭，同时促进了细胞凋亡。值得注意的是，MCF2L-AS1的缺失与顺铂耐药性降低有关。机制上，MCF2L-AS1被确定为miR-874-3p的上游基因，对其表达起负调控作用。在此之后，确定了miR-874-3p的下游靶点STAT3。此外，STAT3的表达与miR-874-3p呈负相关，并受MCF2L-AS1的正调控。一项恢复实验表明，MCF2L-AS1通过结合miR-874-3p和调节STAT3表达来促进LUAD细胞生长。有趣的是，STAT3随后被证实是一种与MCF2L-AS1启动子结合的转录因子，从而增强其转录。