超声微泡介导G3BP1转染对人肝癌HepG2细胞增殖、迁移的影响  

Effects of ultrasound microbubble-mediated G3BP1 transfection on proliferation and migration of human hepatocarcinoma HepG2 Cells

作　　者：黄文凡[1] 周雯 章宜兰[3] 黄炎[4] Huang Wenfan;Zhou Wen;Zhang Yilan;Huang Yan(Department of Ultrasonic,Yuhang District First People’s Hospital,Hangzhou 311100,China;Department of Gastroenterology,Wuhan Yaxin General Hospital,Wuhan 430000,China;Department of Emergency,First Clinical Medical College of Three Gorges University&Yichang Central People’s Hospital,Yichang 443003,China;Department of Pathology,Yuhang District First People’s Hospital,Hangzhou 311100,China)

机构地区：[1]杭州市余杭区第一人民医院超声科,311100 [2]武汉亚心总医院消化内科,430000 [3]三峡大学第一临床医学院&宜昌市中心人民医院急诊科,宜昌443003 [4]杭州市余杭区第一人民医院病理科,311100

出　　处：《中华内分泌外科杂志》2021年第6期622-626,共5页Chinese Journal of Endocrine Surgery

基　　金：浙江省医学会临床科研基金(2019ZYC-A101)。

摘　　要：目的探究超声微泡介导RasGAP SH3结构域结合蛋白1(G3BP1)转染对人肝癌HepG2细胞增殖、迁移的影响。方法超声靶向微泡破坏(ultrasound-targeted microbubble desturction,UTMD)技术介导的沉默G3BP1(si-G3BP1)处理HepG2细胞,通过逆转录-定量聚合酶链反应(RT-qPCR)及Western blot分析G3BP1在HepG2细胞中的表达水平;通过流式细胞术、甲基噻唑蓝(MTT)、EdU染色、集落形成实验、伤口愈合实验、Transwell实验及Western blot分析HepG2细胞增殖和迁移。结果HepG2细胞中沉默G3BP1后,其mRNA和蛋白水平显著降低[(1.01±0.03)vs(0.27±0.03)、(1.02±0.01)vs(0.33±0.04)];UTMD介导的si-G3BP1可明显降低HepG2细胞阳性细胞增殖率[(31.49±3.09)vs(12.51±1.02)]、增殖活力[(1.20±0.13)vs(0.46±0.31)]、EdU阳性细胞率[(99.23±1.01)vs(36.75±4.03)]、集落形成率[(96.45±1.21)vs(32.67±2.62)]、划痕愈合率[(97.58±1.04)vs(42.33±2.56)]、迁移率[(94.28±2.33)vs(39.36±2.51)]及Ki67、细胞周期蛋白D1(Cyclin D1)、波形蛋白(Vimentin)蛋白水平,升高E-钙粘蛋白(E-cadherin)蛋白水平。结论UTMD介导的si-G3BP1可抑制人肝癌HepG2细胞增殖和迁移。Objective To investigate the effects of ultrasound microbubble-mediated RasGAP SH3-binding protein 1(G3BP1)transfection on the proliferation and migration of human liver cancer HepG2 cells.Methods HepG2 cells were treated with ultrasound targeted microbubble destruction(UTMD)technology-mediated si-G3BP1.The expression of G3BP1 in HepG2 cell lines was detected by Western blot,and the silencing efficiency was analyzed by reverse transcription-quantitative polymerase chain reaction(RT-qPCR)and Western blot.HepG2 cell proliferation and migration were analyzed by flow cytometry,methyl thiazolyl tetrazolium(MTT),EdU staining,colony formation experiment,wound healing experiment,Transwell experiment and Western blot.Results After silencing G3BP1 in HepG2 cells,its mRNA and protein levels were significantly reduced(1.01±0.03 vs 0.27±0.03,1.02±0.01 vs 0.33±0.04);UTMD-mediated si-G3BP1 could significantly reduce the proliferation rate(31.49±3.09 vs 12.51±1.02),proliferation activity(1.20±0.13 vs 0.46±0.31),EdU-positive cell rate(99.23±1.01 vs 36.75±4.03),colony formation rate(96.45±1.21 vs 32.67±2.62),scratch healing rate(97.58±1.04 vs 42.33±2.56),migration rate(94.28±2.33 vs 39.36±2.51)and Ki67,Cyclin D1,Vimentin protein levels,increase E-cadherin protein levels.Conclusion UTMD-mediated si-G3BP1 can inhibit the proliferation and migration of human liver cancer HepG2 cells.

关 键 词：RasGAP SH3结构域结合蛋白1 超声靶向微泡破坏 肝癌 HEPG2细胞 增殖 迁移 

分 类 号：R735.7[医药卫生—肿瘤]