Glioblastoma multiforme (GBM) is the most commonly encountered subtype of deadly brain cancer in human adults. It has a high recurrence rate and shows aggressive proliferation. The novel cytotoxic agent temozolomide (TMZ) is now frequently applied as the first-line chemotherapeutic treatment for GBM; however, a considerable number of patients treated with TMZ turn out to be refractory to this drug. Hence, a more effective therapeutic approach is urgently required to overcome this critical issue. Accumulating evidence has shown that both AMPK and AKT are activated by TMZ, while only AMPK contributes to apoptosis via mammalian target of rapamycin (mTOR) inhibition. Accordingly, AKT increases the tumorigenicity and chemoresistance of various tumor cells. In addition, AKT overexpression increases the resistance of glioma cells to TMZ. Cordycepin, a major bioactive component in Cordyceps militaris, exhibits immunomodulatory, anticancer, antioxidant, and anti-inflammatory activities, among other therapeutic effects. To date, whether GBM sensitivity to TMZ can be enhanced by cordycepin largely remains unknown. In the present study, we evaluated the effect of the combined use of cordycepin and TMZ in the treatment of GBM and explored the molecular mechanisms. Notably, we found that treatment with cordycepin led to inhibition of cellular proliferation, migration, and invasion as well as cellular apoptosis and cell cycle arrest in glioma cell lines in vitro. Likewise, the combined treatment with both cordycepin and TMZ synergistically resulted in inhibition of cellular growth, migration, and tumor metastasis as well as induction of cellular apoptosis and cell cycle arrest. Moreover, we also demonstrated that cordycepin effectively enhanced the activation of AMPK and suppressed the activity of AKT, whose activation was only induced by TMZ. Furthermore, there was an apparent reduction in the expression levels of p-mTOR, p-p70S6K, matrix metalloproteinase (MMP)-2, and MMP-9 in the group treated with both cordycepin and TMZ, in comparison with those in the groups treated with either cordycepin or TMZ alone. In vivo, the combination therapy also obviously reduced the tumor volume as well as prolonged the median survival time of xenograft models. In brief, our results suggested that cordycepin augments TMZ sensitivity in human glioma cells at least partially through activation of AMPK and suppression of the AKT signaling pathway. Overall, the combination therapy of cordycepin and TMZ potentially provides a novel option for a better prognosis of patients with GBM in clinical practice.
