Use of Conditioned Media from Electrically‐Stimulated C2C12 Myotubes to Examine the Effect of Muscle Contractile Secretions on Breast Cancer Cell Proliferation

Abstract

Background & PurposeEvidence suggests that secretions following muscle contraction may have benefits regarding breast cancer. This study aimed to examine whether conditioned media from electrically‐stimulated C2C12 myotubes would inhibit proliferation of breast cancer cells in vitro.MethodsMedia collected from electrically‐stimulated C2C12 myotubes was mixed 1:1 with fresh media and applied to either T47D subluminal type breast cancer cells or a K14TRT triple‐negative breast cancer cell line for 24 hours. Breast cancer cell proliferation was measured using the MTT method. In addition, media from electrically‐stimulated and non‐stimulated C2C12 myotubes were subjected to mass spectrometry for comparison.ResultsResults provided an inconsistent picture regarding the approach. When applied at a 1:1 ratio, the conditioned media unexpectedly accelerated the proliferation of T47D cells by more than two‐fold. When applied to K14TRT cells, one experiment demonstrated no effect, while a second showed a robust inhibition of proliferation. The disparate results are likely due to the wide natural variance in proliferation in this cell line. Mass spectrometry analysis did not yield any potential differences in secreted molecules between electrically‐stimulated and non‐stimulated media.ConclusionsThe present results suggest that further work is needed to explore the utility of media from electrically‐stimulated myotubes as a model for muscle contraction‐induced secretions and their effect on breast cancer. Future work will include a dose‐response analysis and the use of ELISA to identify potential molecules.Support or Funding InformationThis study was partially supported by a WSSU Grant from the Office of Science Initiatives. Mass spectrometry was conducted by the Wake Forest Baptist Comprehensive Cancer Center Proteomics and Metabolomics Shared Resource, which is supported by the National Cancer Institute’s Cancer Center Support Grant award number P30CA012197