IR-Surviving NSCLC Cells Exhibit Different Patterns of Molecular and Cellular Reactions Relating to the Multifraction Irradiation Regimen and p53-Family Proteins Expression

Abstract

Radiotherapy is a primary treatment modality for patients with unresectable non-small cell lung cancer (NSCLC). Tumor heterogeneity still poses the central question of cancer radioresistance, whether the presence of a particular cell population inside a tumor undergoing a selective outgrowth during radio- and chemotherapy give rise to metastasis and tumor recurrence. In this study, we examined the impact of two different multifraction X-ray radiation exposure (MFR) regimens, fraction dose escalation (FDE) in the split course and the conventional hypofractionation (HF), on the phenotypic and molecular signatures of four MFR-surviving NSCLC cell sublines derived from parental A549 (p53 wild-type) and H1299 (p53-null) cells, namely A549FR/A549HR, H1299FR/H1299HR cells. We demonstrate that sublines surviving different MFR regimens in a total dose of 60 Gy significantly diverge in their molecular traits related to irradiation regimen and p53 status. The observed changes regarding radiosensitivity, transformation, proliferation, metabolic activity, partial epithelial-to-mesenchymal transition (EMT) program activation and 1D confined migratory behavior (wound healing). For the first time, we demonstrated that MFR exposure led to the significant decrease in the expression of p63 and p73, the p53-family members, in p53null cells, which correlated with the increase in cell polyploidy. We could not find significant differences in FRA1 expression between parental cells and their sublines that survived after any MFR regimen regardless of p53 status. In our study, the FDE regimen probably causes partial EMT program activation in MFR-survived NSCLC cells through either Vimentin upregulation in p53null or an aberrant N-cadherin upregulation in p53wt cells. The HF regimen likely less influences the EMT activation irrespectively of the p53 status of MFR-survived NSCLC cells. Our data highlight that both MFR regimens caused overall higher cell transformation of p53null H1299FR and H1299HR cells than their parental H1299 cells. Moreover, our results indicate that the FDE regimen raised the radioresistance and transformation of MFR-surviving NSCLC cells irrespectively of their p53 status, though the HF regimen demonstrated a similar effect on p53null NSCLC cells only. Our data once again emphasize that NSCLC therapy approaches should become more personalized according to radiation therapy (RT) regimen, tumor histology, and molecular status of critical proteins.

GATA3 Exerts Distinct Transcriptional Functions to Regulate Radiation Resistance in A549 and H1299 Cells

“…It has been demonstrated in the literature that two types of lung adenocarcinoma cells (A549 and H1299) have different sensitivities to radiation, and after conventionally fractionated irradiation regimens, the two types of cells exhibit different apoptosis, metabolic activity, and EMT transformation [ 36 ]. This fully demonstrates that nonsmall cell lung cancer (NSCLC) treatments should become more personalized according to the status of key protein molecules in tumor tissue [ 34 ]. This also coincides with our research.…”

“…This phenomenon may be caused by different cellular genetic backgrounds. A549 cells are p53 wild-type and H1299 cells are p53 deletion-type [32][33][34]. It is well known that p53 is a tumor suppressor and can also regulate the metabolic pathways of cancer cells.…”

Deep Radiotranscriptomics of Non-Small Cell Lung Carcinoma for Assessing Molecular and Histology Subtypes with a Data-Driven Analysis

“…In medicine, the increasing quantities of information could outline the complexity of the underlining biology of specific lesions, especially in oncology. While several efforts have used single source data to investigate and model cancer mechanisms [ 56 , 57 , 58 , 59 , 60 ], our effort is towards the synergistic use of high dimensional and high throughput data (deep features, radiomics and transcriptomics) for identifying the prognostic signatures towards precision decision support in oncology.…”

The Molecular and Cellular Strategies of Glioblastoma and Non-Small-Cell Lung Cancer Cells Conferring Radioresistance

“…Furthermore, it has been observed that the expression of Oct-4, SOX2, and β-catenin proteins markedly increases in adherent H460 cells maintained in a monolayer after IR at a dose of 5 Gy [ 324 ]. Our previous data have suggested that a fraction dose escalation regimen at a total dose of 60 Gy probably causes partial (or hybrid) EMT program activation in multifractionated radiotherapy surviving NSCLC cells through either Vimentin upregulation in p53null or an aberrant N-cadherin upregulation in p53wt cells [ 441 ]. Moreover, we have indicated previously that the hypofractionation regimen IR does not significantly influence horizontal 1D cell migration of multifractionated radiotherapy surviving NSCLC cells, though promoting their migration by 24 h after scratching [ 441 ].…”