[1]燕 婧,冯 芸,李 丹,等.microRNA-185-5p真核表达载体构建及其对胰腺癌Panc-1细胞增殖的影响[J].陕西医学杂志,2021,50(11):1323-1326，1332.[doi:DOI:10.3969/j.issn.1000-7377.2021.11.001]
　YAN Jing,FENG Yun,LI Dan,et al.Construction of microRNA-185-5p overexpression vector and its effect on the proliferation of pancreatic cancer cell Panc-1[J].,2021,50(11):1323-1326，1332.[doi:DOI:10.3969/j.issn.1000-7377.2021.11.001]
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microRNA-185-5p真核表达载体构建及其对胰腺癌Panc-1细胞增殖的影响
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
50
期数:
2021年11期
页码:
1323-1326，1332
栏目:
基础研究
出版日期:
2021-11-05

文章信息/Info

Title:
Construction of microRNA-185-5p overexpression vector and its effect on the proliferation of pancreatic cancer cell Panc-1
作者:
燕 婧; 冯 芸; 李 丹; 高 艺; 张 静
(延安大学医学院,陕西 延安 716000)
Author(s):
YAN Jing; FENG Yun; LI Dan; GAO Yi; ZHANG Jing
(Medical School of Yan'an University,Yan'an 716000,China)
关键词:
miR-185-5p;  真核表达载体;  胰腺癌;  双酶切;  Panc-1细胞;  细胞增殖
Keywords:
miR-185-5p;  Eukaryotic expression vector;  Pancreatic cancer;  Double enzyme digestion;  Panc-1 cells;  Cell proliferation
分类号:
R 735.9
DOI:
DOI:10.3969/j.issn.1000-7377.2021.11.001
文献标志码:
A
摘要:
目的:构建microR-185-5p(miR-185-5p)的过表达载体并研究miR-185-5p对人胰腺癌Panc-1细胞体外增殖的影响。方法:将人工合成的miR-185-5p寡聚核苷酸与经EcoR I和Hind Ⅲ双酶切连接上纯化后的真核表达载体pcDNATM6.2-GW/EmGFP-miR,接着转化进E.Coli Top10,再经过DNA测序、比对检测进行验证; 将构建成功的载体转染进Panc-1细胞中,用倒置荧光显微镜和qRT-PCR方法验证转染效率,用CCK8方法检测miR-185-5p对Panc-1细胞增殖的影响。结果:成功构建了miR-185-5p真核表达载体; 构建好的载体瞬时转染Panc-1细胞后发现miR-185-5p的表达显著增加,并且能够抑制Panc-1细胞的体外增殖能力。结论:miR-185-5p的真核表达载体构建成功,同时证明了miR-185-5p过表达可抑制人胰腺癌细胞Panc-1的增殖。
Abstract:
Objective:To construct microRNA-185-5p(miR-185-5p)overexpression vector,and to explore its effect on the proliferation of human pancreatic cancer cell line Panc-1 in vitro.Methods:The synthetic oligonucleotide of miR-185-5p was combined with the purified eukaryotic expression vector pcDNATM6.2-GW/EmGFP-miR that digested by EcoR Ⅰ and Hind Ⅲ,and then transformed into E.Coli Top10,then verified by DNA sequencing,comparison and detection.The successfully constructed vector was transfected into Panc-1 cells,and the transfection efficiency was determined by inverted fluorescence microscope and qRT-PCR,and the effect of miR-185-5p on the proliferation of Panc-1 cells was detected by CCK8 method.Results:The eukaryotic expression vector of miR-185-5p was successfully constructed.After transient transfection of Panc-1 cells with the constructed vector,the expression of miR-185-5p was significantly increased,and it could inhibit the expression of Panc-1 cells.Conclusion:The eukaryotic expression vector of miR-185-5p is successfully constructed.It is proved that the overexpression of miR-185-5p can inhibit the proliferation of human pancreatic cancer cell Panc-1.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金资助项目(81660492)
更新日期/Last Update: 2021-11-05