Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Oct 15:15:1471996.
doi: 10.3389/fphar.2024.1471996. eCollection 2024.

The single-strand DNA-binding protein SSB1 is involved in the expression of salivary gland radiation injury repair

Xian Wang #  1   2 , Yuetong Wang #  1   2 , Xianglin Zeng #  3 , Haoyu Lu  1   2 , Dongqin Mo  1   2 , Yuetao Li  1   2 , Zhiqing Liu  1   2 , Yude Huang  1   2 , Kun Yu  1 , Daiyou Wang  1   2
Affiliations

The single-strand DNA-binding protein SSB1 is involved in the expression of salivary gland radiation injury repair

Xian Wang et al. Front Pharmacol. .

Abstract

Objectives: Single-strand DNA-binding protein 1 (SSB1) plays a crucial role in the cellular response to DNA damage. This study aimed to explore the expression and regulation of SSB1 in normal rat salivary gland tissues and tissues following radiation, with a specific emphasis on its involvement in the repair of salivary gland injury.

Methods: A total of 45 adult SD rats were randomly assigned to one control group or eight experimental groups. In the control group, five rats were euthanized without irradiation, and their parotid gland tissues were collected for analysis. The experimental groups received a dose of 6 Gy of radiation targeting the head and neck region; subsequently, five rats from each group were euthanized hly to collect parotid gland tissue samples, resulting in a total of eight experimental groups. The expression levels of SSB1, γ-H2AX, and PARP1 in the parotid gland tissues were assessed via immunohistochemistry, while changes in SSB1 gene expression were quantified via RT-qPCR.

Results: No significant morphological differences were observed between the two groups following HE staining. In the immunohistochemistry (IHC) analysis, notable tissue-specific variation in SSB1 expression was identified, with higher levels detected in the ducts than in the acini and connective tissue. The expression of SSB1 gene initially increased post-radiation before subsequently decreasing, ultimately returning to baseline levels, as corroborated by the RT-qPCR results. In contrast, γ-H2AX and PARP1 exhibited minimal expression in the control group; however, their expression peaked at 1 h in the experimental group before gradually declining to levels comparable to those of the control group.

Conclusion: Radiation induces time-dependent upregulation of SSB1 expression in rat salivary glands, indicating that SSB1 may play a role in radiation-induced repair processes.

Keywords: DSB repair; PARP1; radioactive damage; salivary gland; single-strand DNA-binding protein 1; γ-H2AX.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
The function of SSB1 in the repair of DNA double-strand break. SSB1 is integral to the repair process, encompassing the protection of single-stranded DNA, recruitment of the MRN complex, coordination with γ-H2AX and PARP repair proteins to enhance signaling for repair, and regulation of cell cycle arrest mediated by P53 and P21. Additionally, SSB1 amplifies ATM kinase activity, thereby accelerating homologous recombination (HR) and overall DNA damage repair processes, making it a crucial protein for maintaining genomic stability.
FIGURE 2
FIGURE 2
The HE and IHC staining of SSB1 in the parotid gland tissues of the rats. (A–I): Control group and 8 experimental groups, respectively. In the experimental groups, the expression of SSB1 first increased but then decreased over time. The expression was the highest in the 3-h group after radiation (D) and then gradually decreased to the normal level. Except for the 7-h group, there were significant differences between the other experimental groups and the control group (J), P < 0.05.
FIGURE 3
FIGURE 3
IHC staining of γ-H2AX. (A-I) represent the control group and 8 experimental groups, respectively. The positive expression of γ-H2AX was the highest at 1 h (B) and then gradually decreased. Except for the 8-h group, there were significant differences between the other experimental groups and the control group (J), p < 0.05.
FIGURE 4
FIGURE 4
IHC staining of PARP1. (A-I) represent the control group and the 8 experimental groups, respectively. The positive expression of PARP1 peaks at 1 h (B) and subsequently decreases gradually. Within 8 h following radiation exposure, statistically significant differences were observed between the experimental groups and the control group (J), P < 0.05.
FIGURE 5
FIGURE 5
RT-qPCR results of SSB1 mRNA expression. The mRNA expression first increased but then decreased, peaking at 2 h, and there were statistically significant differences compared with that in the control group between 1 and 4 h, P < 0.05.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Adams M. N., Croft L. V., Urquhart A., Saleem M. A. M., Rockstroh A., Duijf P. H. G., et al. (2023). hSSB1 (NABP2/OBFC2B) modulates the DNA damage and androgen-induced transcriptional response in prostate cancer. Prostate 83 (7), 628–640. 10.1002/pros.24496 - DOI - PMC - PubMed
    1. Atorino L., Di Meglio S., Farina B., Jones R., Quesada P. (2001). Rat germinal cells require PARP for repair of DNA damage induced by gamma-irradiation and H2O2 treatment. Eur. J. Cell Biol. 80 (3), 222–229. 10.1078/0171-9335-00153 - DOI - PubMed
    1. Banáth J. P., Klokov D., MacPhail S. H., Banuelos C. A., Olive P. L. (2010). Residual gammaH2AX foci as an indication of lethal DNA lesions. BMC Cancer 10, 4. 10.1186/1471-2407-10-4 - DOI - PMC - PubMed
    1. Benafif S., Hall M. (2015). An update on PARP inhibitors for the treatment of cancer. Onco Targets Ther. 8, 519–528. 10.2147/OTT.S30793 - DOI - PMC - PubMed
    1. Bolderson E., Petermann E., Croft L., Suraweera A., Pandita R. K., Pandita T. K., et al. (2014). Human single-stranded DNA binding protein 1 (hSSB1/NABP2) is required for the stability and repair of stalled replication forks. Nucleic Acids Res. 42 (10), 6326–6336. 10.1093/nar/gku276 - DOI - PMC - PubMed

LinkOut - more resources