PTBP3 mediates TGF-β-induced EMT and metastasis of lung adenocarcinoma

Cell Cycle. 2022 Jul;21(13):1406-1421. doi: 10.1080/15384101.2022.2052530. Epub 2022 Mar 24.

Abstract

Lung adenocarcinoma (LUAD) is associated with a poor prognosis due to early metastasis to distant organs. TGF-β potently induces epithelial-to-mesenchymal transition (EMT) and promotes invasion and metastasis of cancers. However, the mechanisms underlying this alteration are largely unknown. PTBP3 plays a critical role in RNA splicing and transcriptional regulation. Although accumulating evidence has revealed that PTBP3 exhibits a pro-oncogenic role in several cancers, whether and how PTBP3 mediates TGF-β-induced EMT and metastasis in LUAD remains unknown. The expression levels and prognostic value of PTBP3 were analyzed in human LUAD tissues and matched normal tissues. siRNAs and lentivirus-mediated vectors were used to transfect LUAD cell lines. Various in vitro experiments including western blot, qRT-PCR, a luciferase reporter assay, chromatin immunoprecipitation (ChIP), transwell migration and invasion assay and in vivo metastasis experiment were performed to determine the roles of PTBP3 in TGF-β-induced EMT and metastasis. PTBP3 expression was significantly upregulated in patients with LUAD, and high expression of PTBP3 indicated a poor prognosis. Intriguingly, we found that PTBP3 expression level in LUAD cell lines was significantly increased by exogenous TGF-β1 in a Smad-dependent manner. Mechanistically, p-Smad3 was recruited to the PTBP3 promoter and activated its transcription. In turn, PTBP3 knockdown abolished TGF-β1-mediated EMT through the inhibition of Smad2/3 expression. Furthermore, PTBP3 overexpression increased lung and liver metastasis of LUAD cells in vivo. PTBP3 is indispensable to TGF-β-induced EMT and metastasis of LUAD cells and is a novel potential therapeutic target for the treatment of LUAD.

Keywords: EMT; LUAD; PTBP3; TGF-β/Smad signaling; tumor metastasis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma of Lung* / metabolism
  • Adenocarcinoma of Lung* / pathology
  • Cell Line, Tumor
  • Cell Movement / physiology
  • Epithelial-Mesenchymal Transition
  • Humans
  • Lung Neoplasms* / metabolism
  • Lung Neoplasms* / pathology
  • Polypyrimidine Tract-Binding Protein* / genetics
  • Polypyrimidine Tract-Binding Protein* / metabolism
  • Signal Transduction
  • Transforming Growth Factor beta1* / metabolism
  • Transforming Growth Factor beta1* / pharmacology

Substances

  • PTBP3 protein, human
  • Transforming Growth Factor beta1
  • Polypyrimidine Tract-Binding Protein

Grants and funding

This work was supported by the National Natural Science Foundation of China (No.81974053) and Shanghai Committee of Science and Technology (No 18441902300, 16441907800).