Verapamil treatment induces cytoprotective autophagy by modulating cellular metabolism
Verapamil treatment induces cytoprotective autophagy by modulating cellular metabolism
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DOI：
10.1111/febs.14064
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发表时间：
2017-05-01
期刊：
影响因子：
5.4
通讯作者：
Orzechowski, Arkadiusz
中科院分区：
文献类型：
--
作者：
Kania, Elzbieta;Pajak, Beata;Orzechowski, Arkadiusz
维拉帕米是一种l型钙通道阻滞剂，已成功用于治疗心血管疾病。有趣的是，我们最近发现用维拉帕米治疗癌细胞会对自噬产生影响。由于已知自噬可以调节化疗反应，这促使我们更详细地探索维拉帕米对自噬和细胞活力的影响。我们在此报告维拉帕米在一定数量的肿瘤细胞和永生化正常细胞中诱导自噬通量。此外，我们发现，通过氯喹（CQ）处理或CRISPR/ cas9介导的自噬基因Atg7和Atg5的破坏，抑制COLO 205细胞的自噬，导致维拉帕米反应中凋亡标志物的上调。为了寻找这种作用的机制，并且由于自噬通常可以减轻代谢应激，我们研究了维拉帕米对细胞代谢的影响。这表明，在正常前列腺细胞中，维拉帕米降低葡萄糖和糖酵解的中间水平，导致5'-三磷酸腺苷（ATP）耗竭。相反，在COLO 205细胞中，它促进有氧糖酵解和维持ATP。重要的是，我们发现这些细胞的自噬反应与l-乳酸脱氢酶A （LDHA, EC 1.1.1.27）的活性有关，因为LDHA的抑制降低了基础和维拉帕米诱导的自噬，从而降低了细胞活力。总之，这些发现不仅确定了细胞保护性自噬诱导的新机制，而且还强调了维拉帕米联合自噬抑制剂治疗恶性疾病的潜力。
Verapamil, an L-type calcium channel blocker, has been used successfully to treat cardiovascular diseases. Interestingly, we have recently shown that treatment of cancer cells with verapamil causes an effect on autophagy. As autophagy is known to modulate chemotherapy responses, this prompted us to explore the impact of verapamil on autophagy and cell viability in greater detail. We report here that verapamil causes an induction of autophagic flux in a number or tumor cells and immortalized normal cells. Moreover, we found that inhibition of autophagy in COLO 205 cells, via treatment with the chloroquine (CQ) or by CRISPR/Cas9-mediated disruption of the autophagy genes Atg7 and Atg5, causes an upregulation of apoptotic markers in response to verapamil. In search of a mechanism for this effect and because autophagy can often mitigate metabolic stress, we examined the impact of verapamil on cellular metabolism. This revealed that in normal prostate cells, verapamil diminishes glucose and glycolytic intermediate levels leading to adenosine 5'-triphosphate (ATP) depletion. In contrast, in COLO 205 cells it enhances aerobic glycolysis and maintains ATP. Importantly, we found that the autophagic response in these cells is related to the activity of L-lactate dehydrogenase A (LDHA, EC 1.1.1.27), as inhibition of LDHA reduces both basal and verapamil-induced autophagy and consequently decreases cell viability. In summary, these findings not only identify a novel mechanism of cytoprotective autophagy induction but they also highlight the potential of using verapamil together with inhibitors of autophagy for the treatment of malignant disease.