超声微泡介导miR-181c-shRNA质粒转染对人肝癌HepG2细胞增殖、周期和凋亡的影响  

Study on the effects of cell proliferation,cycle and apoptosis of miR-181c-shRNA plasmid transfected into human hepatoma HepG2 cells by microbubble ultrasound contrast agents

作　　者：朱芳方 过新民[2] ZHU Fang-fang;GUO Xin-min(Department of Ultrasound, Foshan Hospital of TCM, Foshan 528000 China;Department of Ultrasound, Guangzhou Red Cross Hospital, Guangzhou 510220 ChinaP<0.05P>0.05P<0.05P<0.05)

机构地区：[1]佛山市中医院超声科,510180 [2]暨南大学附属广州市红会医院超声科,510220

出　　处：《现代消化及介入诊疗》2020年第6期717-723,共7页Modern Interventional Diagnosis and Treatment in Gastroenterology

基　　金：广东省科技发展专项资金(公益研究与能力建设方向)项目(2016A020215015)。

摘　　要：目的探讨miR-181c表达与肝细胞肝癌(HCC)患者预后的相关性及超声微泡介导miR-181c干扰质粒(miR-181c-shRNA)转染对人肝癌HepG2细胞增殖、周期和凋亡的影响。方法下载癌症基因组图谱(TCGA)中HCC患者数据集,分析miR-181c表达与HCC患者生存时间及预后的关系,利用前期实验筛选的最优超声转染条件介导miR-181c-shRNA质粒转染人肝癌HepG2细胞,实时荧光定量PCR(qPCR)检测转染后细胞miR-181c的表达情况,并检测沉默miR-181c表达后细胞增殖、周期及凋亡情况。结果生存分析结果显示,miR-181c高表达患者的无病生存期较低表达患者明显减少,差异有统计学意义(P<0.05),而总生存期相比较差异无统计学意义(P>0.05);Cox比例风险模型提示miR-181c表达是HCC的独立预后因素(P<0.05);转染miR-181c-shRNA质粒后细胞的miR-181c表达下调,人肝癌HepG2细胞的增殖受到抑制、周期受到阻滞(P<0.05),但对凋亡无明显影响。结论miR-181c表达可作为HCC的独立预后因素,超声微泡介导miR-181c-shRNA转染人肝癌HepG2细胞后,miR-181c表达下调,细胞增殖受到抑制、周期发生阻滞,但对凋亡无明显影响,本研究有望为HCC的基因治疗提供新的靶点。Objective To study on the prognostic relationship between the expression of miR-181c with Hepatocellular carcinoma patients and to explore the cell proliferation,cycle and apoptosis of human hepatoma HepG2 cells transfected miR-181c-shRNA plasmid by microbubble ultrasound contrast agents.Methods Analyzed the survival time and prognostic relationship between the expression of miR-181c with HCC patients through the data set of HCC patients which was downloaded in TCGA.MiR-181c-shRNA plasmid was transfected into HepG2 cells of human liver cancer by using the optimal ultrasonic transfection conditions screened in previous experiments.The expression of mir-181c in the transfected cells was detected by real-time fluorescence quantitative PCR(qPCR),and the proliferation,cycle and apoptosis of the cells were further detected after the expression of miR-181c was silenced.Results Survival analysis results indicated that disease-free survival of patients with high mir-181c expression was significantly lower than patients with low expression,and the difference was statistically significant(P<0.05),while the overall survival was not statistically significant(P>0.05).Cox proportional risk model suggested that mir-181c expression was an independent prognostic factor in HCC(P<0.05).Mir-181c expression was down-regulated after transfected with plasmid mir-181c-shrna;the cell proliferation were inhibited,the cell cycle were blocked(P<0.05),while it had no effct on cell apoptosis.Conclusion Mir-181c expression can be used as an independent prognostic factor for HCC.After transfection of human HCC HepG2 cells with mir-181c-shrna mediated by ultrasonic microbubbles,mir-181c expression is down-regulated,which inhibits the cell proliferation,blocksthe cell cycle,but has no effect on cell apoptosis.That may provide a new target for HCC gene therapy.

关 键 词：超声微泡造影剂 miR-181c 基因转染 肝细胞肝癌 

分 类 号：R319[医药卫生—基础医学] R575