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
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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