RESUMEN
AIM: To investigate the changes of proliferation and apoptosis in K562 cells over-expressing miR-10a. METHODS: K562 cells were infected with pAd-pre-miR-10a virus, and then the levels of miR-10a and bcr/abl in transfected K562 cells was detected using RT-PCR and the level of BCR/ABL using Western blotting. The methylthiazolyl tetrazolium (MTT) assay and flow cytometry (FCM) were used to monitor the changes of proliferation and apoptosis of K562 cells after transfection. RESULTS: Compared with the control groups, K562 cells transfected with pAd-pre-miR-10a presented the significantly increased level of miR-10a, the significantly decreased expressions of bcr/abl fusion gene and BCR/ABL fusion protein, the inhibited proliferation ability and the promoted apoptosis (P<0.05). CONCLUSION: The recombinant adenovirus of pAd-pre-miR-10a can inhibit the proliferation of K562 cells and promote cell apoptosis significantly.
Asunto(s)
Adenoviridae/genética , Vectores Genéticos/genética , MicroARNs/genética , Apoptosis/genética , Proliferación Celular , Proteínas de Fusión bcr-abl/genética , Proteínas de Fusión bcr-abl/metabolismo , Expresión Génica , Técnicas de Transferencia de Gen , Humanos , Células K562 , MicroARNs/metabolismoRESUMEN
bcr/abl fusion gene is thought to be a promising target for chronic myelogenous leukemia (CML) patients to enhance immune response after attaining complete remission. In this study, we sought to enhance cellular immunity by co-expression of BCR/ABL and murine IL-12 gene on the tumor cell surface as a glycosyl-phosphatidylinositol (GPI)-form. The successfully constructed plasmid pBudCE4.1-BCR/ABL-GPI-mIL12 resulted in high levels of splenocyte proliferative responses, significant levels of IL-2 and IFNγ, and strong cytotoxic T-lymphocyte (CTL) responses in vitro. In a murine transplant model, the vaccinated mice showed decreased infiltration of leukemia cells and reduced expression of BCR/ABL transcripts and protein in bone marrow cells. Results of the present study indicated that this novel immunization strategy is useful in enhancing immune protection in mice, which would provide new insights into the development of effective vaccines for treating CML.
