[1]许扬梅 龚福生 刘沁颖 刘施佳 黄丽洁 郑秋红.MiR-194过表达和抑制表达对肝癌细胞株Hep-3b中侧群细胞增殖的影响[J].福建医药杂志,2019,41(06):141-144.
　XU Yangmei,GONG Fusheng,LIU Qinying,et al.Influence of miR-194 up and down-expression on liver cancer stem cells[J].FUJIAN MEDICAL JOURNAL,2019,41(06):141-144.
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MiR-194过表达和抑制表达对肝癌细胞株Hep-3b中侧群细胞增殖的影响()
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《福建医药杂志》[ISSN:1002-2600/CN:35-1071/R]

卷:
41
期数:
2019年06期
页码:
141-144
栏目:
基础研究
出版日期:
2019-12-25

文章信息/Info

Title:
Influence of miR-194 up and down-expression on liver cancer stem cells
文章编号:
1002-2600(2019)06-0141-05
作者:
许扬梅 龚福生 刘沁颖 刘施佳 黄丽洁 郑秋红1
福建医科大学附属肿瘤医院 福建省肿瘤医院 福建省肿瘤生物治疗重点实验室(福州 350014)
Author(s):
XU Yangmei;  GONG Fusheng;  LIU Qinying;  LIU Shijia;  HUANG Lijie;  ZHENG Qiuhong.
Fujian Provincial Key Laboratory of Tumor Biotherapy,Fujian Medical University Cancer Hospital& Fujian Cancer Hospital,Fuzhou, Fujian 350014, China
关键词:
微小RNA;  慢病毒载体;  肝癌;  干细胞
Keywords:
MicroRNA;  lentivirus vector;  liver cancer;  stem cell
分类号:
R735.7
文献标志码:
B
摘要:
目的 研究MicroRNA-194(miR-194)过表达和抑制表达对细胞株Hep-3b中侧群细胞增殖功能影响。方法 构建miR-194过表达和抑制表达慢病毒并感染Hep-3b细胞。流式细胞仪鉴定各感染组SP侧群比例,QPCR检测各组细胞miR-194-5p表达,CCK- 8法检测各组细胞增殖能力,软琼脂克隆形成实验检测细胞克隆形成能力。结果 流式细胞仪鉴定发现miR-194过表达Hep-3b组SP细胞含量与miR-194抑制表达Hep-3b组差异具有统计学意义(LSD-t=3.550,P=0.012)。QPCR检测显示miR-194过表达Hep-3b组miR-194-5p水平高于miR-194抑制表达Hep-3b组(LSD-t=4.680,P=0.007)。CCK- 8实验表明miR-194过表达Hep-3b组和miR-194抑制表达Hep-3b组细胞增殖能力差异有统计学意义(LSD-t=5.621,P=0.002)。软琼脂克隆形成实验显示miR-194过表达Hep-3b组比miR-194抑制表达Hep-3b组形成细胞克隆少(LSD-t=5.849,P=0.001)。结论 成功构建了miR-194过表达和抑制细胞株,miR-194抑制组显示明显的干细胞特征,其含有更多SP细胞,具有更强的细胞增殖能力和自我更新能力。
Abstract:
Objective To study the influence of miR-194 up and down-expression Hep-3b cell lines on the proliferation ability of SP cells.Methods Hep-3b was used to construct miR-194 up and down-expression cell lines via lentivirus infection.Flow cytometry was applied to identify SP cells.QPCR assay was adopted to test the expression of miR-194-5p in those cell lines.CCK-8 assay was conducted to study the proliferation ability of cells and soft agar colony formation assay was conducted to test colony-forming ability.Results Each infected cell line maintained good growth.Flow cytometry analysis showed that the SP percentage were significantly different between miR-194 up and down-regulation Hep-3b groups(LSD-t=3.550,P=0.012).QPCR assay of miR-194-5p showed statistic difference between miR-194 up and down-regulation Hep-3b groups(LSD-t=4.680,P=0.007).CCK-8 assay showed different growth ability between miR-194 up and down-regulation Hep-3b groups(LSD-t=5.621,P=0.002).Soft agar colony formation assay results less colonies formed in miR-194 up-regulation Hep-3b group than miR-194 down-regulation Hep-3b group(LSD-t=5.849,P=0.001).Conclusion miR-194 up-regulation Hep-3b group than miR-194 down-regulation Hep-3b group were successfully constructed.miR-194 down-regulation Hep-3b group contains more SP cells, possess stronger growth ability and self-renewal ability, showing stronger proliferation and self-renewal ability.

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更新日期/Last Update: 2019-12-25