Takaoka, S.; Ishii, T.; Umihara, Y.; Otani, R.; Akazawa, S.; Oda, T.; Ogino, Y.; Okino, Y.; Wang, D.-S.; Uchiumi, F. Effect of culture supernatant of Clostridium butyricum TO-A on human DNA repair factor-encoding gene promoters. Preprints2024, 2024100401. https://doi.org/10.20944/preprints202410.0401.v1
APA Style
Takaoka, S., Ishii, T., Umihara, Y., Otani, R., Akazawa, S., Oda, T., Ogino, Y., Okino, Y., Wang, D. S., & Uchiumi, F. (2024). Effect of culture supernatant of Clostridium butyricum TO-A on human DNA repair factor-encoding gene promoters. Preprints. https://doi.org/10.20944/preprints202410.0401.v1
Chicago/Turabian Style
Takaoka, S., Dian-Sheng Wang and Fumiaki Uchiumi. 2024 "Effect of culture supernatant of Clostridium butyricum TO-A on human DNA repair factor-encoding gene promoters" Preprints. https://doi.org/10.20944/preprints202410.0401.v1
Abstract
In this study, Clostridium butyricum TO-A culture supernatant (CBCS) or butyric acid was added to a culture medium of human cervical carcinoma HeLa S3 cells, and changes in DNA repair-related gene promoter activi-ties were investigated. HeLa S3 cells were transfected with a luciferase (Luc) expression vector containing ap-proximately 500-bp of the 5′-upstream region of several human DNA repair-related genes and cultured with a medium containing CBCS (10%) or butyric acid (2.5 mM). Cells were harvested after 19 to 42 h of incubation. Luc assay revealed that the human ATM, PARG, PARP1, and RB1 gene promoter activities were significantly increased. Western blot analysis showed that amounts of the proteins encoded by those genes markedly in-creased. Furthermore, 8, 24, and 48 h after the addition of the CBCS (10%), total RNA was extracted and subjected to RNAseq analysis. The result showed that the expression of several DNA replication/repair-related genes, including NFKB, and the MCM gene groups, decreased markedly after 8 h. However, the expression of the histone genes increased after 24 h. Elucidation of the mechanism by which CBCS and butyrate affect DNA-repair associated protein-encoding gene expression may contribute to the prevention of carcinogenesis, of which the risk rises in accordance with aging.
Medicine and Pharmacology, Pharmacology and Toxicology
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