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Mechanism of MicroRNA-708 Targeting BAMBI in Cell Proliferation, Migration, and
Apoptosis in Mice With Melanoma via the Wnt and TGF-? Signaling Pathways
#MMPMID29466930
Lu HJ
; Yan J
; Jin PY
; Zheng GH
; Zhang HL
; Bai M
; Wu DM
; Lu J
; Zheng YL
Technol Cancer Res Treat
2018[Jan]; 17
(?): 1533034618756784
PMID29466930
show ga
OBJECTIVE: The aim of this study was to evaluate the mechanisms involved with
miRNA-708 and its targeting of bone morphogenetic protein and activin
membrane-bound inhibitor in cell proliferation, migration, and apoptosis in mice
with melanoma via the Wnt and transforming growth factor ? signaling pathways.
METHODS: Sixty mice were recruited of which 40 were subsequently assigned into
the experimental group (22 mice were successfully established as melanoma model
and 18 mice used in tumor xenograft), and the normal control group consisted of
20 mice. B16 cells were assigned to the normal, blank, and negative control,
miR-708 mimics, miR-708 inhibitors, si-BAMBI, and miR-708 inhibitors + si-bone
morphogenetic protein and activin membrane-bound inhibitor groups. Western
blotting and reverse transcription quantitative polymerase chain reaction were
employed to detect the expression levels within the tissues and cell lines. TCF
luciferase reporter (TOP-FLASH) or a control vector (FOP-FLASH) was applied to
detect the activity of the Wnt signaling pathway.
MTT3-(4,5-Dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay, flow
cytometry, scratch test, and Transwell assay were conducted, respectively, for
cell proliferation, apoptosis, migration, and invasion, while tumor xenograft
procedures were performed on the nude mice recruited for the study. RESULTS:
Compared to the normal control group, the model group displayed increased
expressions of bone morphogenetic protein and activin membrane-bound inhibitor,
Wnt10B, P53, and Bcl-2; TOPflash activity; ?-catenin expression; cell
proliferation; migration; and invasion capabilities while decreased expressions
of miR-708, vascular endothelial growth factor, Fas, Bax, Caspase-3, and cleaved
Caspase-3 and apoptosis rate. Compared to the blank and negative control groups,
the miR-708 mimics and small-interfering RNA-bone morphogenetic protein and
activin membrane-bound inhibitor groups exhibited decreases expressions of bone
morphogenetic protein and activin membrane-bound inhibitor, Wnt10B, P53, and
Bcl-2 and decreased proliferation, migration, and invasion capabilities, while
increases in the apoptosis rate, expressions of vascular endothelial growth
factor, Fas, Bax, Caspase-3, and cleaved Caspase-3; however, downregulated levels
of TOPflash activity and ?-catenin expression were recorded. The miR-708
inhibitors group displayed an opposite trend. CONCLUSION: Downregulation of
miR-708-targeted bone morphogenetic protein and activin membrane-bound inhibitor
inhibits the proliferation and migration of melanoma cells through the activation
of the transforming growth factor ? pathway and the suppression of Wnt pathway.
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