Features of senescence associated secretory phenotype (sasp) in gastrointestinal stromal tumors (GIsTs)
- Authors: Ramazanov B.R1, Boichuk S.V1, Rizvanov A.A2
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Affiliations:
- Kazan State Medical University
- Kazan (Volga region) Federal University
- Issue: Vol 9, No 3 (2014)
- Pages: 173-178
- Section: Articles
- URL: https://genescells.ru/2313-1829/article/view/120278
- DOI: https://doi.org/10.23868/gc120278
- ID: 120278
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Abstract
To study the cellular senescence mechanisms and senescence-associated secretory phenotype of the tumor cell lines in response to doxorubicin exposure. The cell lines indicated above were used in present study: human fibroblasts lines BJ, human osteosarcoma cell line U-2 OS and gastrointestinal stromal tumor cell line GIST-T1. Genotoxic stress was induced by the transient exposure of doxorubicin (0.25 ug / ml) for 5 hours. DNA double-strand breaks formation was assessed by an increased expression and focal accumulation of the phosphorylated form of histone 2A (y-H2AX), whereas the activation of DNA repair pathway was assessed by focal accumulation of the phosphorylated form of ATM-kinase (pATM Ser1981). An increase of p-galactosidase activity was estimated by X-gal assay (5-bromo-4-chloro-3-indo-beta-D-galactoside). Interleukin-6 (IL-6) and interleukin-8 (IL-8) production was assessed by ELISA at 3, 6, 9, 12 days after 5h exposure to doxorubicin. Incubation of the tumor cell lines with doxorubicin induced DNA damage which triggered the DNA damage response (DDR) and activated a cellular senescence program associated with typical morphological changes, decrease of Lamin B1 expression and accumulation of p-galactosidase in the cytoplasm known as a traditional senescence markers. We observed a significant increase of IL-6 and IL-8 production in doxorubicin-treated human fibroblasts and tumor U-2 OS cells, whereas GIST-T1 cells secreted IL-6 only. In contrast to non-transformed cells, tumor cells after being exposed to doxorubicin triggered a permanent DNA damage, activation of cellular senescence program and production of proinflammatory cytokines (IL-6 and -8). Senescence-associated secretory phenotype (SASP) in GIST-T1 cells differs from the phenotype observed in the other of cancer cells and normal cells, as well. This might be taken into account during the development of the novel treatment options for the patients with gastrointestinal stromal tumors (GISTs).
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About the authors
B. R Ramazanov
Kazan State Medical University
S. V Boichuk
Kazan State Medical University
A. A Rizvanov
Kazan (Volga region) Federal University
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