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16 May 2019 : Laboratory Research  

Doxorubicin-Induced Cancer Cell Senescence Shows a Time Delay Effect and Is Inhibited by Epithelial-Mesenchymal Transition (EMT)

Xuerui Hu1ABCDEF, Hongqi Zhang2ACDEG*

DOI: 10.12659/MSM.914295

Med Sci Monit 2019; 25:3617-3623

Abstract

BACKGROUND: Senescence is a natural barrier for the body to resist the malignant transformation of its own cells. This work investigated the senescence characteristics of cancer cells in vitro.

MATERIAL AND METHODS: Human cervical cancer HeLa cells were treated with different concentrations of doxorubicin for 3 days, with or without subsequent extended culture in drug-free medium for 6 days. Senescent cell ratios between these 2 culture schemes were calculated. Expression of 2 senescence-associated secretory factors, IL-6 and IL-8, were detected by RT-PCR and ELISA. Doxorubicin treatment induced epithelial-mesenchymal transition in cancer cells. The proportions of senescent cells in epithelial-like and mesenchymal-like sub-groups were calculated. Doxorubicin-treated HeLa cells were stained with Vimentin antibody and sorted by flow cytometry. Senescent cell marker p16ᴵᴺᴷ⁴ᵃ and IL-8 expression in Vimentin-high and Vimentin-low cells were detected by Western blot.

RESULTS: We found that less than 1% of HeLa cells showed senescence phenotype after treatment with doxorubicin for 3 days. However, the proportion of senescent cells was significantly increased when the doxorubicin-treated cells were subsequently cultured in drug-free medium for another 6d. RT-PCR and ELISA results showed that this prolonged culture method could further improve the expression of IL-6 and IL-8. We also found that the senescent cells were mainly epithelial-like type and few presented mesenchymal-like shape. p16ᴵᴺᴷ⁴ᵃ and IL-8 expression were decreased in cell fraction with higher Vimentin expression.

CONCLUSIONS: Our results suggested the existence of time delay effect in doxorubicin-induced senescence of HeLa cells, and epithelial- mesenchymal transition may resist doxorubicin-induced cell senescence.

Keywords: Cell Aging, Hela Cells, Interleukin-6, Interleukin-8, Neoplasms, Time Factors, Vimentin

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750