临床与病理杂志2017，Vol.37Issue(6)：1117-1123,7.DOI:10.3978/j.issn.2095-6959.2017.06.007
超声辐照携带miR-520c-3p和miR-132微泡对体外培养人肝癌细胞Huh7的影响
Effect of ultrasound irradiation carrying miR-520c-3p and miR-132 microbubble on human hepatocellular carcinoma Huh7 cell line cultured in vitro
摘要
Abstract
Objective:To investigate the effect of ultrasound irradiation carrying miR-520c-3p and miR-132 microbubble on human hepatocellular carcinoma Huh7 cell line cultured in vitro.Methods:Human hepatocellular carcinoma Huh7 cell line cultured in vitro was transfected with miR-520c-3p and miR-132 which were carried with ultrasound microbubble.The experiment was divided into 5 groups:control group (group A),miR-520c-3p mimics negative control+miR-132 mimics negative control+microbubble+ultrasound irradiation (group B),miR520c-3p mimics+microbubble+ultrasound irradiation (group C),miR-132 mimics+microbubble+ultrasound irradiation (group D),miR-520c-3p mimics+miR-132 mimics+microbubble+ultrasound irradiation (group E).Ultrasonic irradiation conditions:mechanical index 0.4,irradiation parameters 2.0 w/cm2,continuous irradiation 30 s,interval 60 s,a total of 5 times.The expression of miR-520c-3p,miR-132,phosphatidylinositol-3 (GPC3) and transcription factor YAP mRNA were detected by real-time fluorescence quantitative PCR.The expression of GPC3 and YAP protein was detected by Western blotting.The cell proliferation was detected by flow cytometry.The cell apoptosis was detected by flow cytometry.Results:The relative expression of miR-520c-3p in group C and E was higher than that in group A and B (P＜0.05),but there was no significant change in group D (P＞0.05).The relative expression of miR-132 in group D and E was higher than that in group A and B (P＜0.05),but there was no significant change in group C (P＞0.05).Compared with group A and B,the relative expression of GPC3 protein in group C and E was decreased (P＜0.05),but the expression ofGPC3 mRNA was not changed (P＞0.05).Compared with group A and B,the expression of YAP protein and its mRNA in group C,D and E were decreased (P＜0.05).Compared with group A and B,the proliferation rate of group C,D and E decreased (P＜0.05),and the group E decreased most obviously (P＜0.05).The apoptosis rate of group C,D and E increased (P＜0.05),and the group E apoptosis most obviously (P＜0.05).Conclusion:Ultrasound irradiation carrying miR-520c-3p and miR132 microbubble can significantly inhibit the proliferation of human hepatoma cell line Huh7 and promote its apoptosis,which may be related to the regulation of the expression of GPC3 and YAP.关键词
超声辐照/miR-520c-3p/miR-132/肝细胞癌/磷脂酰肌醇蛋白多糖-3/转录辅助因子YAPKey words
ultrasonic irradiation/miR-520c-3p/miR-132/hepatocellular carcinoma/glypican-3/YAP-associated protein引用本文复制引用
潘青云,雷长江,李媛,胡福荣,李磊,黄剑彬,曾诚..超声辐照携带miR-520c-3p和miR-132微泡对体外培养人肝癌细胞Huh7的影响[J].临床与病理杂志,2017,37(6):1117-1123,7.基金项目
湖北省自然科学基金(2016CFB591) （2016CFB591）
武汉市科技局应用基础研究(2015061701011630) （2015061701011630）
第四批汉阳知音英才计划.This work was supported by the Natural Science Foundation of Hubei Province (2016CFB591) （2016CFB591）
Applied Fundamental Research Project of Wuhan Municipal Science and Technology Bureau (2015061701011630) and the Fourth Batch of"Hanyang Talent Associate Program",China. （2015061701011630）
