RESUMEN

MicroRNAs (miRNA), as a class of non-coding RNAs, play a crucial role in normal hematopoiesis. Recent studies on miRNA profiling during hematopoiesis showed miR-10a down-regulation in megakaryocytic differentiation. Here we investigated whether decrease in miR-10a can differentiate umbilical cord blood CD133+ cells to megakaryocytic series. We ectopically induced miR-10a down-regulation by locked nucleic acid anti-miR-10a transfection of CD133+ cells. The megakaryocytic differentiation was evaluated by CD42/CD61 and CD41 surface expression and colonogenic capacity in Megacult media. In addition, real-time polymerase chain reaction was done for detection of miR-10a and its target HoxA1 mRNA. HOXA1 protein expression was detected by flow cytometry as mean fluorescent intensity that shows the antibody reaction proteins. Our findings showed megakaryocytic differentiation of about 28% of umbilical cord blood CD133+ stem cells with bright expression of CD42/CD61 and CD41 in parallel with increase in HoxA1 mRNA and protein level. Colony forming of CD61+ cells in Megacult demonstrated the colonogenic capacity of differentiated progenitor cells. In conclusion, MiR-10a has a role in megakaryocyte differentiation of stem cells via HOXA1 transcription factor targeting.

Asunto(s)

Diferenciación Celular/genética , Regulación hacia Abajo , Células Madre Hematopoyéticas/metabolismo , Megacariocitos/metabolismo , MicroARNs/genética , Antígeno AC133 , Antígenos CD/metabolismo , Células Cultivadas , Sangre Fetal/citología , Citometría de Flujo , Expresión Génica , Técnicas de Silenciamiento del Gen , Glicoproteínas/metabolismo , Células Madre Hematopoyéticas/citología , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Humanos , Integrina beta3/metabolismo , Megacariocitos/citología , Péptidos/metabolismo , Complejo GPIb-IX de Glicoproteína Plaquetaria/metabolismo , Glicoproteína IIb de Membrana Plaquetaria/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Transcripción/genética , Factores de Transcripción/metabolismo