Abstract
Connexins are proteins that form gap junctions. Perturbations in the cell membrane reportedly promote changes in the expression profile of connexins. Electroporation promotes destabilization by applying electrical pulses, and this procedure is used in electrochemotherapy and gene therapy, among others. This in vitro work aimed to study the interference of electroporation on the expression profile of GJB2 (Cx26 gene) and Connexin 26 in melanoma cell line B16/BL6. The techniques of immunocytochemistry, Western blot, and real-time PCR were used. After electroporation, cells showed a transient decrease in GJB2 mRNA. The immunostaining of Cx26 showed no noticeable change after electroporation at different time points. However, Western blot showed a significant reduction in Cx26 30 min after electroporation. Our results showed that electroporation interferes transiently in the expression of Connexin 26 in melanoma and are consistent with the idea that electroporation is a process of intense stress that promotes cell homeostatic imbalance and results in disruption of cell physiological processes such as transcription and translation.
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Acknowledgments
This study is part of the PhD thesis of Marcelo Monte-Mor Rangel in the Experimental and Comparative Pathology Program of the School of Veterinary Medicine and Animal Science of the University of Sao Paulo, Brazil. Marcelo Monte Mor Rangel was the recipient of a fellowship from Fundação de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP, Proc.2008/55534-1). This study has been supported by Grants from FAPESP (Proc. 2007/59391-8).
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Rangel, M.M.M., Chaible, L.M., Nagamine, M.K. et al. Electroporation Transiently Decreases GJB2 (Connexin 26) Expression in B16/BL6 Melanoma Cell Line. J Membrane Biol 248, 47–52 (2015). https://doi.org/10.1007/s00232-014-9735-z
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DOI: https://doi.org/10.1007/s00232-014-9735-z