The Differentiation of C2C12 Cells to Myotube by Paraquat

The Differentiation of C2C12 Cells to Myotube by Paraquat,10.1007/978-90-481-3892-0_35,Masaaki Okabe,Koichi Akiyama,Sogo Nishimoto,Takuya Sugahara,Yos

The Differentiation of C2C12 Cells to Myotube by Paraquat  
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Paraquat (1,1’-dimethyl-4,4’-bipyridinium dichloride), producing reactive oxygen species (ROS) to show its toxicity is one of herbicides once widely used. Although the toxicity of paraquat on human and animals is too strong, paraquat is still in use in many developing countries. Therefore it is important to know the risk of paraquat on mammals. Effect of paraquat on the muscle of mammals is not well investigated, but it has been observed that interstitial cells differentiated into muscle cells in the lung of paraquat-injected monkey (Fukuda et al., Am J Pathol, 118(3): 452–475). Here we studied the effect of paraquat on the differentiation of mouse myoblast cell line C2C12 cells to myotube. C2C12 cells were maintained in Dulbecco’s modified eagle medium containing 10% fetal bovine serum at 37°C, 5% CO2. Paraquat (0.1 ˜ 1000 ng/mL) was added to the medium and C2C12 cells were cultured in each medium for 12 days. Then the cells were harvested and applied to SDS-PAGE and western-blotting with anti-myosin heavy chain (MCH) antibody. Light microscope observation revealed that C2C12 cells morphologically changed to myotube depending on the concentration of paraquat even in undifferentiating condition. The amount of MCH, the molecular marker of muscle differentiation, increased depending on the concentration of paraquat. Paraquat produces ROS to show its toxicity. We also studied the effect of ascorbic acid on paraquat-dependent differentiation of C2C12 cells. C2C12 cells were cultured with paraquat and ascorbic acids in the same conditions stated above. As a result it was observed that ascorbic acid (10˜100 μM) did not inhibit the differentiation of C2C12 cells, and the overproduction of MCH by paraquat was not decreased by the ascorbic acid. Furthermore, the existence of H2O2 (0.1˜100 μM) did not induce the differentiation of C2C12 cells. These results suggest that the differentiation of C2C12 cells to myotube is modulated by paraquat in ROS-independent manner.
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