Long non-coding RNA NEAT1 promotes bone metastasis of prostate cancer through N6-methyladenosine

Abstract

Background N6-methyladenosine (m6A) is the most prevalent messenger RNA modification in mammalian cells. However, the disease relevant function of m6A on specific oncogenic long non-coding RNAs (ncRNAs) is not well understood. Methods We analyzed the m6A status using patients samples and bone metastatic PDXs. Through m6A high-throughput sequencing, we identified the m6A sites on NEAT1–1 in prostate bone metastatic PDXs. Mass spec assay showed interaction among NEAT1–1, CYCLINL1 and CDK19. RNA EMSA, RNA pull-down, mutagenesis, CLIP, western blot, ChIP and ChIRP assays were used to investigate the molecular mechanisms underlying the functions of m6A on NEAT1–1. Loss-of function and rescued experiments were executed to detect the biological roles of m6A on NEAT1–1 in the PDX cell phenotypes in vivo. Results In this study, we identified 4 credible m6A sites on long ncRNA NEAT1–1. High m6A level of NEAT1–1 was related to bone metastasis of prostate cancer and m6A level of NEAT1–1 was a powerful predictor of eventual death. Transcribed NEAT1–1 served as a bridge to facility the binding between CYCLINL1 and CDK19 and promoted the Pol II ser2 phosphorylation. Importantly, depletion of NEAT1–1or decreased m6A of NEAT1–1 impaired Pol II Ser-2p level in the promoter of RUNX2. Overexpression of NEAT1–1 induced cancer cell metastasis to lung and bone; xenograft growth and shortened the survival of mice, but NEAT1–1 with m6A site mutation failed to do these. Conclusion Collectively, the findings indicate that m6A on ncRNA NEAT1–1 takes critical role in regulating Pol II ser2 phosphorylation and may be novel specific target for bone metastasis cancer therapy and diagnosis. New complex CYCLINL1/CDK19/NEAT1–1 might provide new insight into the potential mechanism of the pathogenesis and development of bone metastatic prostate cancer.

Analysis of m6A RNA Methylation-Related Genes in Liver Hepatocellular Carcinoma and Their Correlation with Survival

“…In previous studies, m6A modification of RNA were proved to play an important role in tumorigenesis and metastasis. Wen et al found that m6A on ncRNA NEAT1-1 plays a critical role in the pathogenesis and development of bone metastatic prostate cancer [ 8 ]. Chang et al demonstrated that overexpression of YTH domain-containing family protein 3 (YTHDF3) could induce the transcription of m6A-enriched genes to promote breast cancer metastasis [ 9 ].…”

N6-methyladenosine-dependent signalling in cancer progression and insights into cancer therapies

“…In addition, modification by m6A can promote the degradation of carcinogenic lncRNA XIST through the YTHDF2-dependent pathways and inhibit the proliferation and metastasis of colorectal cancer [ 145 ]. m6A modification can also stabilize LncNEAT1, which acts as a bridge between CYCLINL1 and CDK19, and promote the phosphorylation of Pol II ser2, leading to bone metastasis of prostate cancer [ 146 ]. LINC00460 directly interacts with IGF2BP2 and DHX9 to assist the recognition protein IGF2BP2 to recognize and stabilize HMGA1 mRNA, enhance the protein expression of HMGA1, and promote the proliferation and metastasis of colorectal cancer [ 147 ].…”

Role of RNA N6-Methyladenosine Modification in Male Infertility and Genital System Tumors

“…All the above analysis methods and R package were implemented by R foundation for statistical computing (2020) version 4.0.3 and software packages ggplot2 and pheatmap. MELLT3 regulates the expression of Integrin β1 (ITGB1) and a high m6A level of LncRNA NEAT1-1 promotes the bone and lung metastasis of PCa Wen et al, 2020). Du and Li et al showed that YTHDF2 performed Oncogene functions in PCa through the KDM5A/miRNA-495/YTHDF2/m6A-MOB3B axis and the downstream AKT phosphorylation of METTL3/YTHDF2/LHPP/NKX 3-1, respectively (Du et al, 2020;.…”

“…In 2020, several studies have shown that m6A writer MELLT3 and VIRMA serve an oncogenic role to promotes the development and progression of PCa by Wnt pathway ( Ma et al, 2020 ) and promoting the translation of MYC ( Yuan et al, 2020 ), lncRNAs CCAT1/2 ( Barros-Silva et al, 2020 ). Other studies have proposed that MELLT3 regulates the expression of Integrin β1 (ITGB1) and a high m6A level of LncRNA NEAT1–1 promotes the bone and lung metastasis of PCa ( Li E. et al, 2020 ; Wen et al, 2020 ). Du and Li et al showed that YTHDF2 performed Oncogene functions in PCa through the KDM5A/miRNA-495/YTHDF2/m6A-MOB3B axis and the downstream AKT phosphorylation of METTL3/YTHDF2/LHPP/NKX 3-1, respectively ( Du et al, 2020 ; Li J. et al, 2020 ).…”