Journal Article

LncRNA PKMYT1AR promotes cancer stem cell maintenance in non-small cell lung cancer via activating Wnt signaling pathway

Yuan, Yixiao, Zhao, Song, Jiang, Xiulin, Xiong, Qiuxia, He, Yaomei, Duan, Lincan, Liu, Peishen, Chen, Yongbin, Jiang, Liping, Shen, Qiushuo, Yang, Cuiping

2021

Overview

Background Non-small cell lung cancer (NSCLC) is the most common type of human lung cancers, which has diverse pathological features. Although many signaling pathways and therapeutic targets have been defined to play important roles in NSCLC, limiting efficacies have been achieved. Methods Bioinformatics methods were used to identify differential long non-coding RNA expression in NSCLC. Real-time RT-PCR experiments were used to examine the expression pattern of lncRNA PKMYT1AR, miR-485-5p. Both in vitro and in vivo functional assays were performed to investigate the functional role of PKMYT1AR/miR-485-5p/PKMYT1 axis on regulating cell proliferation, migration and tumor growth. Dual luciferase reporter assay, fluorescent in situ hybridization (FISH), immunoblot, co-immunoprecipitation experiments were used to verify the molecular mechanism. Result Here, we identify a human-specific long non-coding RNA (lncRNA, ENST00000595422), termed PKMYT1AR (PKMYT1 associated lncRNA), that is induced in NSCLC by Yin Yang 1 (YY1) factor, especially in cancerous cell lines (H358, H1975, H1299, H1650, A549 and SPC-A1) compared to that in normal human bronchial epithelium cell line (BEAS-2B). We show that PKMYT1AR high expression correlates with worse clinical outcome, and knockdown of PKMYT1AR inhibits tumor cell proliferation, migration and xenograft tumor formation abilities. Bioinformatic analysis and a luciferase assay demonstrate that PKMYT1AR directly interacts with miR-485-5p to attenuate the inhibitory role on its downstream oncogenic factor PKMYT1 (the protein kinase, membrane-associated tyrosine/threonine 1) in NSCLC. Furthermore, we uncover that miR-485-5p is downregulated in both cancerous cell lines and peripheral blood serum isolated from NSCLC patients compared to reciprocal control groups. Consistently, forced expression of miR-485-5p inhibits the proliferation and migration abilities of tumor cells. Moreover, we provide evidence showing that PKMYT1AR targeting antisense oligonucleotide (ASO) dramatically inhibit tumor growth in vivo. Mechanistic study shows that PKMYT1AR/ miR-485-5p /PKMYT1 axis promotes cancer stem cells (CSCs) maintenance in NSCLC via inhibiting β-TrCP1 mediated ubiquitin degradation of β-catenin proteins, which in turn causes enhanced tumorigenesis. Conclusions Our findings reveal the critical role of PKMYT1AR/miR-485-5p /PKMYT1 axis during NSCLC progression, which could be used as novel therapeutic targets in the future.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: 3' Untranslated Regions / Analysis / Animals / Antisense oligonucleotides / Apoptosis / Bioinformatics / Biological products / Biomedical and Life Sciences / Biomedicine / Cancer Research / Cancer stem cells (CSCs) / Cancer therapies / Carcinoma, Non-Small-Cell Lung - etiology / Carcinoma, Non-Small-Cell Lung - metabolism / Carcinoma, Non-Small-Cell Lung - mortality / Carcinoma, Non-Small-Cell Lung - pathology / Cell adhesion & migration / Cell culture / Cell cycle / Cell growth / Cell Line, Tumor / Cell migration / Cell Movement - genetics / Cell proliferation / Cell Proliferation - genetics / Cloning / Cytogenetics / Drug resistance / Epithelium / Fluorescence in situ hybridization / Gene expression / Gene Expression Regulation, Neoplastic / Health aspects / Humans / Immunoprecipitation / Kinases / Luciferase / Lung cancer / Lung cancer, Non-small cell / Lung cancer, Small cell / Lung Neoplasms - etiology / Lung Neoplasms - metabolism / Lung Neoplasms - mortality / Lung Neoplasms - pathology / Male / Medical prognosis / Membrane proteins / Membrane Proteins - antagonists & inhibitors / Membrane Proteins - genetics / Mice / MicroRNAs / miR-485-5p / Molecular Targeted Therapy / Neoplastic Stem Cells - metabolism / Non-coding RNA / Non-small cell lung cancer / Non-small cell lung carcinoma / Oligonucleotides, Antisense / Oncology / Peripheral blood / PKMYT1 / PKMYT1AR / Polymerase chain reaction / Prognosis / Protein Binding / Protein kinase / Protein kinases / Protein Serine-Threonine Kinases - antagonists & inhibitors / Protein Serine-Threonine Kinases - genetics / Protein Stability / Protein-Tyrosine Kinases - antagonists & inhibitors / Protein-Tyrosine Kinases - genetics / RNA / RNA Interference / RNA, Long Noncoding - genetics / Signal transduction / Small cell lung carcinoma / Stem cells / Therapeutic applications / Threonine / Tumor cells / Tumorigenesis / Tyrosine / Ubiquitin / Vectors (Biology) / Wnt protein / Wnt Signaling Pathway / Xenografts