Imatinib and Dasatinib Provoke Mitochondrial Dysfunction Leading to Oxidative Stress in C2C12 Myotubes and Human RD Cells
Imatinib and Dasatinib Provoke Mitochondrial Dysfunction Leading to Oxidative Stress in C2C12 Myotubes and Human RD Cells
复制标题
DOI：
10.3389/fphar.2020.01106
复制
发表时间：
2020-07-23
影响因子：
5.6
通讯作者：
Kraehenbuehl, Stephan
中科院分区：
文献类型：
--
作者：
Bouitbir, Jamal;Panajatovic, Miljenko Valentin;Kraehenbuehl, Stephan
酪氨酸激酶抑制剂（TKIs）可引起患者骨骼肌毒性，但其潜在机制尚不清楚。当前研究的目的是更好地表征线粒体在tki相关肌毒性中的作用。我们将C2C12小鼠成肌细胞和肌管以及人横纹肌肉瘤细胞（RD细胞）暴露于伊马替尼（1-100 μ M）、厄洛替尼（1-20 μ M）和达沙替尼（0.001-100 μ M）中24小时。在C2C12成肌细胞中，伊马替尼在50 μ M时具有膜毒性，并在20 μ M时耗尽细胞ATP库。在C2C12肌管中，暴露于伊马替尼的ATP在50 μ M时开始耗尽，但未检测到膜毒性。在达沙替尼作用下的成肌细胞和肌管中，膜毒性分别在0.5 μ M和2 μ M时开始，ATP分别在0.1 μ M和0.2 μ M时明显下降。当RD细胞暴露于伊马替尼时，ATP在20 μ M时开始耗竭，而膜毒性未检测到。达沙替尼在20 μ M时具有膜毒性，在0.5 μ M时已耗尽细胞ATP库。厄洛替尼在两种细胞模型中均无毒性。伊马替尼（20 μ M）和达沙替尼（1 μ M）降低了两种细胞模型中复合物I的活性。此外，肌管中两种TKIs的线粒体膜电位（δ psi m）均被耗散。在RD细胞中，只有达沙替尼能降低δ psi m。两种TKIs均增加了两种细胞系的线粒体超氧化物积累，并降低了线粒体拷贝数。因此，它们增加了超氧化物歧化酶（SOD） 2和硫氧还蛋白2的蛋白表达以及caspase 3的裂解，表明C2C12肌管发生凋亡。此外，在两种细胞模型中，当RD细胞暴露于达沙替尼时，sod1和sod2的mRNA表达均增加。此外，达沙替尼增加了atrogin1和murf-1的mRNA表达，这是参与肌肉萎缩的重要转录因子。在暴露于伊马替尼的RD细胞中，atroggin -1的mRNA表达也增加。综上所述，伊马替尼和达沙替尼对小鼠C2C12肌管和人RD细胞具有线粒体毒性。这两种TKIs诱导的线粒体超氧化物积累是由于复合物I的抑制，可能与线粒体和肌细胞增殖受损有关。
Tyrosine kinase inhibitors (TKIs) can cause skeletal muscle toxicity in patients, but the underlying mechanisms are mostly unclear. The goal of the current study was to better characterize the role of mitochondria in TKI-associated myotoxicity. We exposed C2C12 murine myoblasts and myotubes as well as human rhabdomyosarcoma cells (RD cells) for 24 h to imatinib (1-100 mu M), erlotinib (1-20 mu M), and dasatinib (0.001-100 mu M). In C2C12 myoblasts, imatinib was membrane toxic at 50 mu M and depleted the cellular ATP pool at 20 mu M. In C2C12 myotubes exposed to imatinib, ATP depletion started at 50 mu M whereas membrane toxicity was not detectable. In myoblasts and myotubes exposed to dasatinib, membrane toxicity started at 0.5 mu M and 2 mu M, respectively, and the ATP drop was visible at 0.1 mu M and 0.2 mu M, respectively. When RD cells were exposed to imatinib, ATP depletion started at 20 mu M whereas membrane toxicity was not detectable. Dasatinib was membrane toxic at 20 mu M and depleted the cellular ATP pool already at 0.5 mu M. Erlotinib was not toxic in both cell models. Imatinib (20 mu M) and dasatinib (1 mu M) reduced complex I activity in both cell models. Moreover, the mitochondrial membrane potential (Delta psi m) was dissipated for both TKIs in myotubes. In RD cells, the Delta psi m was reduced only by dasatinib. Both TKIs increased mitochondrial superoxide accumulation and decreased the mitochondrial copy number in both cell lines. In consequence, they increased protein expression of superoxide dismutase (SOD) 2 and thioredoxin 2 and cleavage of caspase 3, indicating apoptosis in C2C12 myotubes. Moreover, in both cell models, the mRNA expression ofSod1andSod2increased when RD cells were exposed to dasatinib. Furthermore, dasatinib increased the mRNA expression ofatrogin-1andmurf-1, which are important transcription factors involved in muscle atrophy. The mRNA expression ofatrogin-1increased also in RD cells exposed to imatinib. In conclusion, imatinib and dasatinib are mitochondrial toxicants in mouse C2C12 myotubes and human RD cells. Mitochondrial superoxide accumulation induced by these two TKIs is due to the inhibition of complex I and is probably related to impaired mitochondrial and myocyte proliferation.