LNCAROD is stabilized by m6A methylation and promotes cancer progression via forming a ternary complex with HSPA1A and YBX1 in head and neck squamous cell carcinoma

Abstract

Head and neck squamous cell carcinoma (HNSCC) constitute approximately 4% of all cancers worldwide. In this study, we analyzed the expression profile of the long noncoding RNA (lncRNA) of 502 HNSCC patients from The Cancer Genome Atlas database. Among the differentially expressed lncRNAs between HNSCC and normal samples, LNCAROD is overexpressed in HNSCC and associated with advanced T stage and shortened overall survival. The N6-methyladenosine (m6A) modification mediated by METTL3 and METTL14 enhanced the stability of LNCAROD in HNSCC cells. Depletion of LNCAROD attenuated cell proliferation, mobility in vitro, and tumorigenicity in vivo, whereas overexpression of LNCAROD exerted opposite effects. LNCAROD is mainly distributed in nucleus and binds with YBX1 and HSPA1A proteins. Silencing either YBX1 or HSPA1A did not affect the level of LNCAROD. However, loss of LNCAROD led to shortened half-life of YBX1 protein. Mechanistically, LNCAROD protected YBX1 from proteasomal degradation by facilitating YBX1-HSPA1A protein-protein interaction. Depletion of HSPA1A in LNCAROD-overexpressing cells resulted in accelerated proteasomal degradation of YBX1 protein.Moreover, re-expression of Flag-YBX1 in LNCAROD-silenced cells rescued malignant behavior of HNSCC cells. Our study indicates that LNCAROD is an oncogenic lncRNA and dysregulation of m6A modification might account for aberrant expression of LNCAROD in HNSCC. LNCAROD acts as a scaffold for the interaction between YBX1 and HSPA1A, preventing proteasomal degradation of YBX1 in HNSCC cells.Abbreviations ASO, antisense oligonucleotide; CCK-8, cell counting kit-8; HNSCC, head and neck squamous cell carcinoma; HSPA1A, heat-shock 70-kDa protein 1A; LNCAROD, lncRNA-activating regulator of DKK1; LncRNA, long noncoding RNA; m6A, N6-methyladenosine; METTL14, methyltransferase-like 14; METTL3, methyltransferase-like 3; OS, overall survival; RIP, RNA immunoprecipitation; TSCC, tongue squamous cell carcinoma; YBX1, Y box binding protein 1.

Novel insights into the interplay between m6A modification and noncoding RNAs in cancer

“…Long non-coding RNA 00958 (LINC00958) is upregulated by METTL3 and facilitates HCC cell migration and invasion by sponging miR-3619-5p [65]. METTL3/14 enhance the migration of head and neck squamous cell carcinoma (HNSCC) by upregulating lncRNA activating regulator of DKK1 (LNCAROD) [66]. METTL3-family with sequence similarity 225 member A (FAM225A)-integrin β3 (ITGB3)-FAK/PI3K/Akt axis facilitates the metastasis of nasopharyngeal cancer [67].…”

Long non‑coding RNA expression profiles and related regulatory networks in areca nut chewing‑induced tongue squamous cell carcinoma

“…lncRNAs are non-coding RNA transcripts that exceed 200 nucleotides, and regulate gene expression at the epigenetic, transcriptional and post-transcriptional levels (55). Increasing evidence has demonstrated that aberrant lncRNA expression and dysfunctional lncRNA activity are associated with tumor initiation and progression (56)(57)(58)(59)(60)(61)(62)(63)(64). However, a full transcriptome analysis of the interactions between mRNAs and lncRNAs, and related regulatory networks in areca nut chewing induced TSCC have not yet been reported.…”

Jack of all trades? The versatility of RNA in DNA double-strand break repair

“…LncRNA LNCAROD, which is up-regulated in HNSCC, is highly m 6 A modified by METTL3 and METTL14 in these samples. m 6 A modification stabilises LNCAROD, and LNCAROD expression correlates with poor prognosis and tumorigenicity in HNSCC [ 128 ]. In gastric cancer, m 6 A-associated genes were found to be dysregulated, and poor prognosis correlated with high expression of FTO and WTAP [ 129 ].…”