Busulfan inhibits growth of human osteosarcoma through miR-200 family microRNAs in vitro and in vivo.
Bottom Line: Here, we showed that busulfan dose-dependently reduced the cell viability and proliferation, and induced cell apoptosis, senescence, and reactive oxygen species levels in two osteosarcoma cell lines.Moreover, a series of loss-of-function and gain-of-function experiments further indicated that busulfan may have its anti-osteosarcoma effect by upregulating the microRNA-200 (miR-200) family which subsequently downregulated its target genes ZEB1 and ZEB2.Taken together, our data suggest that busulfan may have an anti-osteosarcoma effect through downregulating ZEB1 and ZEB2 through activating the miR-200 family, highlighting a possibility of using busulfan as a novel therapy for osteosarcoma.
Affiliation: 169th Hospital, School of Medicine, Hunan Normal University, Changsha, China.Show MeSH
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Mentions: To define whether busulfan produced its anti-osteosarcoma effect through downregulation of the miRNA-200 family and by modulation of expression of its downstream targets ZEB1 and ZEB2, we inhibited the expression of miR-200a, miR-200b, miR-200c, miR-141, and miR-429, or overexpressed ZEB1 and ZEB1, by miRNA antisense oligonucleotide transfection or plasmid transfection (p-ZEB1 or p-ZEB2), respectively. We found that downregulation of all five members in the miR-200 family reversed the effect induced by busulfan in terms of cell viability, proliferation, level of ROS apoptosis, and cell senescence (Fig. 3a). Similarly, we found that upregulation of ZEB1 or ZEB2 expression also reversed the anti-osteosarcoma effect of busulfan (Fig. 3b), without affecting the expression levels of the miR-200 family (data not shown). Of note, downregulation of all five members in the miR-200 family resulted in upregulation of ZEB1 and ZEB2 (Fig. 3c). Taken together, these loss-of-function experiments thus indicate that busulfan may have its anti-osteosarcoma effect by upregulating ZEB1 and ZEB2 through activating the miRNA-200 family.
Affiliation: 169th Hospital, School of Medicine, Hunan Normal University, Changsha, China.