Low and high LET radiation‐induced apoptosis in M059J and M059K cells

Abstract

Purpose: To investigate and compare the ability of DNA‐dependent protein kinase (DNA‐PK)‐deficient and ‐proficient cells to undergo apoptosis after exposure to low and high linear energy transfer (LET) radiation.

Materials and methods: A human glioma cell line M059J lacking the catalytic subunit of DNA‐PK (DNA‐PKcs) and its DNA‐PKcs‐proficient counterpart, M059K, were exposed to 1 and 4 Gy of accelerated nitrogen ions (¹⁴N, 140 eV nm⁻¹, 8–12 Gy min⁻¹) or ⁶⁰Co γ‐rays (0.2 eV nm⁻¹, 0.7 Gy min⁻¹). The induction of apoptosis was studied up to 144 h post‐irradiation using two different methods: morphological characterization of apoptotic cells after fluorescent staining and cell size distribution analysis to detect apoptotic bodies. In parallel, protein expression of DNA‐PKcs and poly(ADP‐ribose) polymerase (PARP) as well as DNA‐PK and caspase‐3 activity were investigated.

Results: Low and high LET radiations (4 Gy) induced a time‐dependent apoptotic response in both cell lines. Low LET radiation induced a significantly elevated apoptotic response in M059J as compared with M059K cells at 144 h post‐irradiation. Following high LET radiation exposure, there was no difference between the cell lines at this time. PARP cleavage was detected in M059J cells following both low and high LET irradiation, while only high LET radiation induced PARP cleavage in M059K cells. These cleavages occurred in the absence of caspase‐3 activation.

Conclusions: M059J and M059K cells both display radiation‐induced apoptosis, which occur independently of caspase‐3 activation. The apoptotic course differs between the two cell lines and is dependent on the quality of radiation.