The CD44high Subpopulation of Multifraction Irradiation-Surviving NSCLC Cells Exhibits Partial EMT-Program Activation and DNA Damage Response Depending on Their p53 Status
Autor: | Anna Chigasova, Taisia Blokhina, Lina Alhaddad, Nadezhda Smetanina, Margarita Pustovalova, Sergey B. Leonov, Andreyan N. Osipov, Ilmira Gilmutdinova, Roman N. Chuprov-Netochin, Petr Eremin |
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Jazyk: | angličtina |
Rok vydání: | 2021 |
Předmět: |
cancer stem cells
p53 DNA End-Joining Repair Lung Neoplasms Cell Radiation Tolerance lcsh:Chemistry Cell Movement Carcinoma Non-Small-Cell Lung DNA Breaks Double-Stranded lcsh:QH301-705.5 Spectroscopy education.field_of_study biology General Medicine Computer Science Applications radioresistance medicine.anatomical_structure epithelial-to-mesenchymal transition Epithelial-Mesenchymal Transition Population Article Catalysis Inorganic Chemistry Cancer stem cell Cell Line Tumor Radioresistance Autophagy medicine Humans Epithelial–mesenchymal transition Radiosensitivity Physical and Theoretical Chemistry education Molecular Biology non-small cell lung cancer X-Rays Organic Chemistry CD44 Recombinational DNA Repair DNA lcsh:Biology (General) lcsh:QD1-999 A549 Cells Cancer cell biology.protein Cancer research Rad51 Rad51 Recombinase Tumor Suppressor Protein p53 |
Zdroj: | International Journal of Molecular Sciences Volume 22 Issue 5 International Journal of Molecular Sciences, Vol 22, Iss 2369, p 2369 (2021) |
ISSN: | 1422-0067 |
DOI: | 10.3390/ijms22052369 |
Popis: | Ionizing radiation (IR) is used for patients diagnosed with unresectable non-small cell lung cancer (NSCLC). However, radiotherapy remains largely palliative due to the survival of specific cell subpopulations. In the present study, the sublines of NSCLC cells, A549IR (p53wt) and H1299IR (p53null) survived multifraction X-ray radiation exposure (MFR) at a total dose of 60 Gy were investigated three weeks after the MFR course. We compared radiosensitivity (colony formation), expression of epithelial-mesenchymal transition (EMT) markers, migration activity, autophagy, and HR-dependent DNA double-strand break (DSB) repair in the bulk and entire CD44high/CD166high CSC-like populations of both parental and MFR survived NSCLC cells. We demonstrated that the p53 status affected: the pattern of expression of N-cadherin, E-cadherin, Vimentin, witnessing the appearance of EMT-like phenotype of MFR-surviving sublines 1D confined migratory behavior (wound healing) the capability of an irradiated cell to continue to divide and form a colony of NSCLC cells before and after MFR influencing the CD44/CD166 expression level in MFR-surviving NSCLC cells after additional single irradiation. Our data further emphasize the impact of p53 status on the decay of γH2AX foci and the associated efficacy of the DSB repair in NSCLC cells survived after MFR. We revealed that Rad51 protein might play a principal role in MFR-surviving of p53 null NSCLC cells promoting DNA DSB repair by homologous recombination (HR) pathway. The proportion of Rad51 + cells elevated in CD44high/CD166high population in MFR-surviving p53wt and p53null sublines and their parental cells. The p53wt ensures DNA-PK-mediated DSB repair for both parental and MFR-surviving cells irrespectively of a subsequent additional single irradiation. Whereas in the absence of p53, a dose-dependent increase of DNA-PK-mediated non-homologous end joining (NHEJ) occurred as an early post-irradiation response is more intensive in the CSC-like population MFR-surviving H1299IR, compared to their parental H1299 cells. Our study strictly observed a significantly higher content of LC3 + cells in the CD44high/CD166high populations of p53wt MFR-surviving cells, which enriched the CSC-like cells in contrast to their p53null counterparts. The additional 2 Gy and 5 Gy X-ray exposure leads to the dose-dependent increase in the proportion of LC3 + cells in CD44high/CD166high population of both parental p53wt and p53null, but not MFR-surviving NSCLC sublines. Our data indicated that autophagy is not necessarily associated with CSC-like cells’ radiosensitivity, emphasizing that careful assessment of other milestone processes (such as senescence and autophagy-p53-Zeb1 axis) of primary radiation responses may provide new potential targets modulated for therapeutic benefit through radiosensitizing cancer cells while rescuing normal tissue. Our findings also shed light on the intricate crosstalk between autophagy and the p53-related EMT, by which MFR-surviving cells might obtain an invasive phenotype and metastatic potential. |
Databáze: | OpenAIRE |
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