Abstract:

Objective　To identify the phenotype of miR-148a-3p in regulating angiogenesis. Methods　The primary

tissues of miR-148a knock-out mice were extracted and immunohistochemical staining was used to investigate the effect of

miR-148a knockout on angiogenesis. By means of miRNA Mimic transfection, miR-148a-3p was overexpressed in human

umbilical vein endothelial cells (hUVECs). Thereafter, the influence of miR-148a-3p overexpression on angiogenesis was

studied in aspects of Matrigel-based tube forming assay, cell proliferation, cell migration, qPCR, etc. The effects of miR-148a-

3p on angiogenesis were clarified in terms of knock-out and overexpression. Results　Compared with heterozygous littermates,

the miR-148a knock-out mice had an abnormal distribution of type-H vessels in the bone tissue. The ex-vivo fetal metatarsal

experiments further confirmed the enhanced angiogenesis ability in absence of miR-148a. The expression level of miR-148a-

3p was overexpressed by miR-148a-3p mimic in vascular endothelial cells. It was found that the overexpression of miR-148a-

3p did not affect the proliferation ability of hUVECs, but could interfere with angiogenesis by inhibiting its migration ability, and

affect the formation of lamellipodia and tubes. Conclusion　miR-148a-3p can inhibit angiogenesis both in-vivo and in-vitro,

including sprouting, migration, and tube formation.

Key words:

miR-148a-3p, 　Angiogenesis, 　Gene knock-out mice, 　Phenotype identification, 　Type H vessels