Efeito do laser de baixa intensidade sobre o perfil transcricional de RNAm em mioblastos C2C12
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Data
2018-03-05
Autores
Ferreira, Juarez Henrique
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Universidade Estadual Paulista (Unesp)
Resumo
A irradiação pelo laser de baixa intensidade (LBI) tem sido utilizada como um método nãoinvasivo para promover ou acelerar a capacidade de regeneração muscular. No entanto, os mecanismos moleculares regulatórios pelos quais o LBI exerce esses efeitos, permanecem em grande parte desconhecidos. Nosso objetivo foi realizar uma análise de sequenciamento de RNA (RNA-Seq) em mioblastos C2C12 após LBI. Foram realizadas as taxas de viabilidade, migração, proliferação e os dados de RNA-Seq dos mioblastos C2C12, identificando 514 genes diferencialmente expressos após LBI. Em seguida, uma análise de ontologia genética e das vias dos genes diferencialmente expressos revelaram transcritos relacionadas ao ciclo celular, biogênese ribossômica, resposta ao estresse, migração celular, estrutura morfológica e proliferação de células musculares. Após, cruzamos nossos dados de RNA-Seq com dados de transcriptomas disponíveis em base de dados públicas, com dados de diferenciação miogênica que mostraram um total de 42 transcritos sobrepostos (mioblastos vs miotubos). Este conjunto de transcritos compartilhados mostrou que os mioblastos irradiados pelo LBI, possuem um perfil transcricional semelhante ao de miotubo, agrupando-se distante do perfil transcricional dos mioblastos. Concluíndo, revelamos pela primeira vez que LBI, induz a uma expressão de um grande conjunto de RNAm, que codificam proteínas reguladoras do ciclo celular que podem controlar a proliferação e diferenciação de mioblastos em miotubos. Importantemente, esse conjunto de RNAm, revelou um perfil transcricional semelhante ao dos miotubos, fornencendo novos conhecimentos para a compreensão das alterações moleculares específicas subjacentes aos efeitos da irradiação por LBI em células do músculo esquelético.
Low-level laser irradiation (LLLT) has been used as a non-invasive method to promote or accelerate muscular regeneration capability. However, the regulatory molecular mechanisms by which LLLT exerts these effects remain largely unknown. Our goal was to perform a RNAsequencing (RNA-Seq) analysis in C2C12 myoblasts after LLLT. C2C12 myoblasts viability, migration, proliferation and RNA-Seq were performed, identifying 514 differentially expressed genes after LLLT. Next, gene ontology and pathway analysis of the differentially expressed genes revealed transcripts among categories related to cell cycle, ribosome biogenesis, response to stress, cell migration, morphological structure and muscle cell proliferation. After, we intersected our RNA-Seq data with transcriptomes publicly available myogenic differentiation data that showed a total of 42 overlapping transcripts (myoblasts vs myotube). This set of shared transcripts showed that the LLLT-myoblasts have a myotube-like profile, clustering away from the myoblast profile. In conclusion, we revealed for the first time that LLLT induces the expression a large set of mRNAs encoding for cell cycle regulatory proteins that may control myoblasts proliferation and differentiation into myotubes. Importantly, these set of mRNA revealed a myotube-like transcriptional profile and provided new insights to the understanding of the specific molecular changes underlying the effects of LLLT irradiation on skeletal muscle cells.
Low-level laser irradiation (LLLT) has been used as a non-invasive method to promote or accelerate muscular regeneration capability. However, the regulatory molecular mechanisms by which LLLT exerts these effects remain largely unknown. Our goal was to perform a RNAsequencing (RNA-Seq) analysis in C2C12 myoblasts after LLLT. C2C12 myoblasts viability, migration, proliferation and RNA-Seq were performed, identifying 514 differentially expressed genes after LLLT. Next, gene ontology and pathway analysis of the differentially expressed genes revealed transcripts among categories related to cell cycle, ribosome biogenesis, response to stress, cell migration, morphological structure and muscle cell proliferation. After, we intersected our RNA-Seq data with transcriptomes publicly available myogenic differentiation data that showed a total of 42 overlapping transcripts (myoblasts vs myotube). This set of shared transcripts showed that the LLLT-myoblasts have a myotube-like profile, clustering away from the myoblast profile. In conclusion, we revealed for the first time that LLLT induces the expression a large set of mRNAs encoding for cell cycle regulatory proteins that may control myoblasts proliferation and differentiation into myotubes. Importantly, these set of mRNA revealed a myotube-like transcriptional profile and provided new insights to the understanding of the specific molecular changes underlying the effects of LLLT irradiation on skeletal muscle cells.
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transcriptoma, terapia com laser, desenvolvimento muscular, músculo esquelético, crescimento muscular, ciclo celular, transcriptome, laser treatment, muscular development, skeletal muscle, muscle growth, cell cycle