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Different effects of omega-3 fatty acids on the cell cycle in C2C12 myoblast proliferation

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Abstract

Polyunsaturated fatty acids (PUFAs) are important molecules for human health. We investigated the effects of three major omega-3 PUFAs on C2C12 myoblast proliferation. Both docosahexaenoic (DHA) and eicosapentaenoic (EPA) acids decreased cell growth, whereas linolenic (ALA) acid did not, compared with the control. Cell cycle analysis showed that G1 phase duration was increased markedly and S-phase duration was decreased by DHA and EPA. In contrast, there was no change in the G1 or S-phase duration when the cells were treated with linolenic acid. To determine how DHA and EPA affected the cell cycle, cyclins and MAPK proteins were investigated. Western blotting and real-time quantitative PCR showed that DHA and EPA decreased cyclin E and CDK2 levels at both the protein and mRNA level. Also, MAPK phosphorylation levels were decreased by treatment with DHA and EPA. Our results indicated that different kinds of n-3 PUFA differentially affected myoblast cell proliferation. DHA and EPA decreased skeletal muscle cell proliferation through a mechanism involving MAPK-ERK.

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Acknowledgments

The study was supported by the earmarked fund for Modern Agro-industry Technology Research Systems of China (CARAS-37), and National Transgenic Breeding Project of China (2011ZX08009-003-006).

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Correspondence to Ning Li.

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Fig. A1

Effect of fatty acids on the proliferation of C2C12 cells. DHA and EPA (10 μM) were added to cells. Cell proliferation curves are shown for 120 h of culture. The viability of cells was determined using the MTT assay. Data represent the means ± SEM of at least five separate experiments, each performed in six wells. (▲) DHA 22:6n-3; (■) EPA 20:5n-3; (◆) ALA 18:3n-3; (×) Control (TIFF 719 kb)

Fig. A2

Cell cycle of DHA-arrested cells changed back to normal after changed the growth medium. (A) cell cycle of DHA-arrested cells; (B) cell cycle of DHA-arrested cells with normal growth medium for 24 h. Results were from at least three repeated experiment. Values are means ± SEM. *, *** P < 0.05 and 0.001, respectively (TIFF 271 kb)

Fig. A3

Effects of Omega-3 fatty acids on C2C12 myoblasts cell viability. Cells were treated with different concentrations (50 μM and 100 μM) of fatty acids for 72 h. Viability was quantitated by Trypan blue exclusion method. The number of viable cells is expressed as a percentage. Black column were treated with 50 μM related fatty acids, white column were treated with 100 μM related fatty acids, Control group were treated with equivalent volume of the ethanol. Results are mean ± SEM of at least five separate experiments. There were no significant differences between control and fatty acids-treated groups (TIFF 585 kb)

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Peng, Y., Zheng, Y., Zhang, Y. et al. Different effects of omega-3 fatty acids on the cell cycle in C2C12 myoblast proliferation. Mol Cell Biochem 367, 165–173 (2012). https://doi.org/10.1007/s11010-012-1329-4

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