SP3-induced Timeless transcription contributes to cell growth of lung adenocarcinoma cells

Ping Tian; Dajun Du; Li Yang; Nan Zhou; Ling Tao

doi:10.1371/journal.pone.0298295

SP3-induced Timeless transcription contributes to cell growth of lung adenocarcinoma cells

PLoS One. 2024 Feb 14;19(2):e0298295. doi: 10.1371/journal.pone.0298295. eCollection 2024.

Authors

Ping Tian¹, Dajun Du², Li Yang³, Nan Zhou⁴, Ling Tao³

Affiliations

¹ Medical School, Xinyang Vocational and Technical College, Xinyang, Henan, China.
² Department of Surgical Oncology, Xinyang Central Hospital, Xinyang, Henan, China.
³ Inspection School, Xinyang Vocational and Technical College, Xinyang, Henan, China.
⁴ Department of Medical Oncology, Xinyang Central Hospital, Xinyang, Henan, China.

Abstract

Background: Timeless is well-known for its key role in replication checkpoints. Recent studies reveal the involvement of Timeless and specificity protein (SP) 1 in human malignancies. However, no evidence proved the interaction between SP3 and Timeless in lung adenocarcinoma (LUAD).

Methods: The expression and clinical significance of Timeless were analyzed using the LUAD dataset downloaded from the Cancer Genome Atlas (TCGA). Lentivirus-mediated Timeless knockdown in A549 cells was used to examine the role of Timeless in cell proliferation and pemetrexed (PEM) resistance. Transcription factors (TFs) bound to the Timeless promoter were identified by DNA pull-down technology with HPLC-MS/MS analysis and analyzed by the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. Dual-luciferase reporter assay was used to determine the activity of SP3 in Timeless transcription.

Results: Timeless was overexpressed in LUAD samples, and it could serve as a potential diagnostic or prognostic biomarker for LUAD patients. shTimeless-mediated knockdown of Timeless reduced cell viability and proliferation and sensitized PEM-resistant A549 cells to PEM. Four fragments (F1: 1-373 bp), (F2: 374-962 bp), (F4: 1274-1645 bp), and (F5: 1646-2000bp) were confirmed as the TF binding profiles of the Timeless promoter. KEGG analysis showed that the TFs bound to the Timeless promoter had relevance to spliceosome, RNA transport, and mRNA surveillance pathways. SP3 promoted the transcription of Timeless via the F2 fragment (374-962 bp) binding motif.

Conclusion: Upregulation of Timeless mediated by SP3 promotes LUAD cell proliferation, providing evidence to support that targeting the SP3/Timeless axis may be a potential therapeutic strategy against LUAD.

Copyright: © 2024 Tian et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

MeSH terms

Adenocarcinoma of Lung* / pathology
Cell Line, Tumor
Cell Proliferation / genetics
Gene Expression Regulation, Neoplastic
Humans
Lung Neoplasms* / pathology
Tandem Mass Spectrometry

Substances

TIMELESS protein, human
SP3 protein, human

Grants and funding

• L T is Initials of the authors who received each award, • There is no grant number awarded to each author, • the full name of each funder is Science and technology research projects in Henan Province: Effect and mechanism of cryptotanshinone enhancing chemotherapeutic Pemetrexed on non-small cell lung cancer (No. 222102310520). • There is no URL of each funder website, • NO- The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.