肿瘤药学2019，Vol.9Issue(5)：736-743,8.DOI:10.3969/j.issn.2095-1264.2019.05.07
miR-133a靶向EGFL7抑制肝癌血管新生的实验研究★
MicroRNA-133a Exerts Anti-angiogenetic Effect in Hepatocellular Carcinoma by Downregulating EGFL7★
摘要
Abstract
Objective To investigate the targeted relationship between miR-133a and epidermal growth factor-like domain 7 (EGFL7), as well as the possible influence of such targeted relationship on the hepatocellular carcinoma (HCC) angiogenesis. Methods Luciferase reporter gene assay was performed to determine the relationship between miR-133a and EGFL7. HCC cell line QGY-7703 was cultured in vitro , and there were 5 groups of transfected cell lines, including blank, miR-133a mimic, miR-133a mimic control, miR-133a inhibitor and miR-133a inhibitor control groups. Then, reverse transcription quantitative polymerase chain reaction (RT-PCR) and Western blotting were used to detect expression of miR-133a, EGFL7 and VEGF. MTT assay was conducted for cell viability. The tumor formation assay was implemented by inject-ing the transfected cells of the blank, miR-133a mimic and miR-133a inhibitor groups into BALB/c rats, with tumor volume and microvessel density measured. The automatic biochemical analyzer was applied to determine the alanine aminotransferase (ALT) level. ELISA and immuno-histochemistry were carried out to detect the VEGF level. Results The luciferase reporter gene assay showed miR-133a can block the expres-sion of the EGFL7. The transfection of miR-133a mimic and miR-133a inhibitor showed that EGFL7 mRNA and protein expressions, VEGF expression and cell viability evidently declined, as the miR-133a expression increased (P<0.05). The tumor formation assay indicated that mice treated with miR-133a mimic had the smallest tumor volume, while mice treated with miR-133a inhibitor had the largest tumor volume (P<0.01). The decline of miR-133a expression could lead to an increase of ALT level and VEGF expression in serum (P<0.05), as well as an increase of MVD and VEGF expression in transplanted tumor tissue(P<0.05). Overexpression of miR-133a decreased the ALT level and VEGF expression in serum (P<0.05), and the VEGF expression and MVD in transplanted tumor tissue (P<0.05). Conclusion Our data demonstrates that miR-133a reduces tumor cells proliferation, MVD and angiogenesis in HCC by downregulating EGFL7.关键词
microRNA/miR-133a/EGFL7/肝癌/肿瘤/血管生成/VEGF/微血管密度Key words
microRNA-133a/Epidermal growth factor-like domain 7/Hepatocellular carcinoma/Angiogenesis/Vascular endothelial growth factor/Microvessel density分类
医药卫生引用本文复制引用
沈二栋,翁洁,文芳,肖佳,罗盘,谢王踢,粟钰淇,黄辉云..miR-133a靶向EGFL7抑制肝癌血管新生的实验研究★[J].肿瘤药学,2019,9(5):736-743,8.基金项目
湖南省自然科学基金面上项目(2017JJ2260). （2017JJ2260）
