The lncRNA MIR99AHG directs alternative splicing of SMARCA1 by PTBP1 to enable invadopodia formation in colorectal cancer cells

Sci Signal. 2023 Sep 19;16(803):eadh4210. doi: 10.1126/scisignal.adh4210. Epub 2023 Sep 19.

Abstract

Alternative splicing regulates gene expression and functional diversity and is often dysregulated in human cancers. Here, we discovered that the long noncoding RNA (lncRNA) MIR99AHG regulated alternative splicing to alter the activity of a chromatin remodeler and promote metastatic behaviors in colorectal cancer (CRC). MIR99AHG was abundant in invasive CRC cells and metastatic tumors from patients and promoted motility and invasion in cultured CRC cells. MIR99AHG bound to and stabilized the RNA splicing factor PTBP1, and this complex increased cassette exon inclusion in the mRNA encoding the chromatin remodeling gene SMARCA1. Specifically, MIR99AHG altered the nature of PTBP1 binding to the splice sites on intron 12 of SMARCA1 pre-mRNA, thereby triggering a splicing switch from skipping to including exon 13 to produce the long isoform, SMARCA1-L. SMARCA1, but not SMARCA1-L, suppressed invadopodia formation, cell migration, and invasion. Analysis of CRC samples revealed that the abundance of MIR99AHG transcript positively correlated with that of SMARCA1-L mRNA and PTBP1 protein and with poor prognosis in patients with CRC. Furthermore, TGF-β1 secretion from cancer-associated fibroblasts increased MIR99AHG expression in CRC cells. Our findings identify an lncRNA that is induced by cues from the tumor microenvironment and that interacts with PTBP1 to regulate alternative splicing, potentially providing a therapeutic target and predictive biomarker for metastatic CRC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing
  • Chromatin
  • Colorectal Neoplasms* / genetics
  • Heterogeneous-Nuclear Ribonucleoproteins / genetics
  • Humans
  • Podosomes*
  • Polypyrimidine Tract-Binding Protein / genetics
  • RNA Splicing
  • RNA, Long Noncoding* / genetics
  • Tumor Microenvironment

Substances

  • Chromatin
  • Heterogeneous-Nuclear Ribonucleoproteins
  • Polypyrimidine Tract-Binding Protein
  • PTBP1 protein, human
  • RNA, Long Noncoding
  • SMARCA1 protein, human
  • MIRN99 microRNA, human