Data from: Gal-1 promotes lung cancer cell survival by enhancing PARP1/H1.2 interaction to promote DNA repair upon DNA damage response
Data files
May 23, 2025 version files 210.33 MB
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Mass_spectrometry_data.xlsx
3.01 MB
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README.md
4 KB
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Transcriptome_data.gz
207.32 MB
Abstract
Galectin-1 (Gal-1), a member of the galectin family, has emerged as a regulator of tumor progression. Several studies have reported the up-regulation of Gal-1 expression in multiple cancer cells and its promotion on tumor proliferation. However, the mechanism by which Gal-1 promotes tumor growth remains to be thoroughly understood. In this study, it was discovered that high expression of Gal-1 in viral cancers was inversely correlated with the overall survival of patients. Through constructing Gal-1-overexpressing cell lines, it was uncovered that cell proliferation and colony formation were significantly improved. The results of transcriptomic and proximity-labeling-based proteomic analyses indicated that Gal-1 interacted with PARP1 and histone H1.2 in lung cancer cells. In the case of etoposide treatment leading to DNA double-strand breaks, Gal-1 accelerated the degradation of H1.2 by enhancing its interaction with PARP1 and promoting its PARylation modification. It caused the activation of downstream DNA repair pathways such as the ATM and NBS1 signaling pathways, thus reducing apoptosis. Moreover, Gal-1 inhibitors TDG and OTX008 could restore cell sensitivity to etoposide. This study provides new clues for the role of Gal-1 in the development of tumors and renders suggestions for the treatment of patients with high Gal-1 expression in the clinic.
Dataset DOI: 10.5061/dryad.wstqjq2wg
Description of the data and file structure
This dataset encompasses transcriptomic data and proximity labeling-based mass spectrometry (MS) data derived from experimental analyses. For the transcriptomic profiling, we performed RNA sequencing on four experimental groups: the parental A549 cell line, the genetically engineered A549-GAL-1 cell line, and both cell lines treated with 100 μM Etoposide for 2 hours. Comparative analysis revealed a panel of differentially transcribed genes (adjusted p-value < 0.05, fold-change > 2) for subsequent functional investigations. Regarding the proximity labeling proteomics, 293T-GAL-1-TurboID stable cells underwent biotinylation treatment followed by streptavidin-mediated magnetic bead enrichment. The processed samples were subjected to LC-MS/MS analysis at the Core Facility of Life Sciences College, Wuhan University, identifying 1,718 putative GAL-1-interacting proteins through stringent filtering criteria. Integrative multi-omics analysis demonstrated significant enrichment of DNA damage repair pathway components in both differentially expressed gene clusters and GAL-1-associated protein networks, suggesting GAL-1's pivotal role in genomic stability maintenance.
Files and variables
File: Mass_spectrometry_data.xlsx
Description: The provided table enables systematic extraction of key proteomic parameters, including: Protein Accession (unique identifier), Description (functional annotation), q-value (false discovery rate-adjusted significance metric), Sum PEP Score (posterior error probability aggregation reflecting identification confidence), Sequence Coverage (percentage of amino acid residues detected by peptide-spectrum matches), Total Peptides (number of distinct peptide sequences mapped), PSMs (spectral counts supporting peptide identification), Unique Peptides (proteotypic sequences unambiguously assigned to the protein), Amino Acid Length (AAs), Molecular Weight (MW), Isoelectric Point (pI), Post-Translational Modifications (PTMs identified with localization probabilities), and Peptide Sequences (exact matched proteolytic fragments). NA indicate missing values.
File: Transcriptome_data.gz
Description: Total RNA was extracted from four experimental groups: parental A549 cells, genetically modified A549-GAL-1 cells, and both cell lines post-treatment with the therapeutic agent. The RNA samples were subjected to high-throughput sequencing at BGI Genomics (Shenzhen, China). Raw sequencing data underwent rigorous quality control through SOAPnuke (v2.2.1) with the following parameters: 1) Removal of adapter-contaminated reads (≥5 bp overlap), 2) Exclusion of low-quality sequences (>40% bases with Phred score < 20), and 3) Elimination of reads with >5% N content. Processed reads were subsequently aligned to the human reference genome (GRCh38) using HISAT2 (v2.2.1) with default splicing-aware parameters, achieving an average alignment rate of 92.4% ± 1.3% across samples. Transcript quantification was performed through RSEM (v1.3.1) employing the expectation-maximization algorithm, generating normalized expression values (TPM and FPKM) with batch effect correction using Combat-seq. This analytical pipeline ensured the reliability of downstream differential expression analysis through DESeq2 .
Code/software
Microsoft Excel can be used to view Mass spectrometry data.
The raw date data were filtered using SOAPnuke (v2.2.1) software for mRNA alignment, and HISTA2 (v2.2.1) software was used to align the filtered data to the human genome
Access information
Other publicly accessible locations of the data:
- The data that support the findings of this study are openly available in Dryad.
Proximity-labeling MS and analysis
The stable Galectin-1-TurboID-NLS-Flag A549 cells were incubated with 500μM biotin， 5 mM MgCl2, and 1 mM ATP for 12h. The cells were harvested and washed, then lysed with the IP lysis buffer. The lysate was incubated with blocked streptavidin beads(Smart-Lifesciences,#SM017K01) with a rotator at 4℃ overnight. Then, wash the beads following the kit`s protocols. 10% of the beads, which were eluted by 1×SDS loading buffer and boiled for 10 mins, were used for to silver stain assay and Immunoblotting. The rest 90% of the beads were sent to the Wuhan University instrument and equipment sharing platform for MS analysis.
RNA seq and analysis
Total RNA was extracted from cells with the indicated treatment and sent to BGI to construct a single-stranded circular DNA library. The constructed library is qualitatively inspected, and sequencing is carried out after it is qualified.