Accumulation of Temozolomide-Induced Apoptosis, Senescence and DNA Damage by Metronomic Dose Schedule: A Proof-of-Principle Study with Glioblastoma Cells

SIMPLE SUMMARY: Severe toxic side effects do not allow unlimited dose escalation of anticancer drugs, and the doses used in cancer therapy are therefore often rather low regarding the required target concentration. For temozolomide (TMZ), which is used in glioblastoma therapy, single high dose protocols are used in nearly all experimental studies, while the drug is administered repeatedly on patients, with a daily (metronomic) low dose schedule. Here, we show that the therapeutically relevant glioblastoma cell death and senescence responses do accumulate if a high dose of TMZ is split up in small low doses. The data support the metronomic dose schedule and suggest that even low doses are effective in glioblastoma therapy. The predominance and accumulation of TMZ-refractory senescent survivors may provide an explanation for the overall low curative response. ABSTRACT: Temozolomide (TMZ), a first-line drug in glioma therapy, targets the tumor DNA at various sites. One of the DNA alkylation products is O(6)-methylguanine (O(6)MeG), which is, in the low dose range of TMZ, responsible for nearly all genotoxic and cytotoxic effects relevant for cancer therapy. There is, however, a dispute regarding whether the TMZ concentration in the tumor tissue in patients is sufficient to elicit a significant cytotoxic or cytostatic response. Although treatment with TMZ occurs repeatedly with daily doses (metronomic dose schedule) and in view of the short half-life of the drug it is unclear whether doses are accumulating. Here, we addressed the question whether repeated low doses elicit similar effects in glioblastoma cells than a high cumulative dose. We show that repeated treatments with a low dose of TMZ (5 × 5 µM) caused an accumulation of cytotoxicity through apoptosis, cytostasis through cellular senescence, and DNA double-strand breaks, which was similar to the responses induced by a single cumulative dose of 25 µM TMZ. This finding, together with the previously reported linear dose–response curves, support the notion that TMZ is able to trigger a significant cytotoxic and cytostatic effect in vivo if the low-dose metronomic schedule is applied.