王杰;喻超;周显飞;朱昌豪;黄巍;江建新;
WANG Jie;YU Chao;ZHOU Xianfei;ZHU Changhao;HUANG Wei;JIANG Jianxin;Department of Hepatic-Biliary-Pancreatic Surgery,the Affiliated Hospital of Guizhou Medical University;

• 1:贵州医科大学附院肝胆外科

摘要(Abstract)：

目的:探讨miRNA-1181(miR-1181)对人胰腺癌细胞增殖能力的影响。方法:采用实时荧光定量聚合酶链反应(qRT-RCR)检测miR-1181在4株人胰腺癌细胞系及1株人正常胰腺上皮细胞系中的表达,选取miR-1181表达差异较明显的PANC-1和MIA PaCa-2细胞系进行后续实验;构建miR-1181过表达慢病毒载体(miR-1181U)以及空载慢病毒载体(NC),采用qRT-PCR检测感染效率;在体外采用CCK-8实验,平板克隆实验检测miR-1181对胰腺癌细胞增殖能力的影响;建立裸鼠皮下成瘤模型,用免疫组化实验检测miR-1181对胰腺癌细胞成瘤能力的影响。结果:qRT-PCR显示4株胰腺癌细胞系中miR-1181表达量低于正常胰腺上皮细胞(P<0.05);过表达miR-1181后PANC-1和MIA PaCa-2细胞增殖能力明显受到抑制(P<0.05),细胞集落形成数量明显减少(P<0.05);裸鼠皮下成瘤模型显示过表达miR-1181后肿瘤体积及免疫组化指标Ki67均明显低于对照组(P<0.05)。结论:过表达的miR-1181能在体内体外抑制胰腺癌细胞的增殖,有望成为胰腺癌治疗的新靶点。
Objective:To investigate the effect of microRNA-1181 on the proliferation of pancreatic cancer.Methods:Quantitative polymerase chain reaction(qRT-RCR) was adopted to detect the expression of miR-1181 in 4 human pancreatic cancer cell lines and 1 human normal pancreatic epithelial cell lines.The PANC-1 and MIA PaCa-2 cell lines with significantly different miR-1181 expression were selected for subsequent experiments.The miR-1181 over-expression lentiviral vector(miR-1181U) and the empty lentiviral vector vector(NC) were constructed and qRT-PCR was used to detect infection efficiency.The effect of miR-1181 on the proliferation of pancreatic cancer cells was detected by CCK-8 assay and Plate cloning experiment in vitro.The subcutaneous tumor model of nude mice was established,and the effect of miR-1181 on the ability of pancreatic cancer cells was detected by immunohistochemistry.Results:qRT-PCR showed that the expression of miR-1181 in 4 pancreatic cancer cell lines was significantly lower than that in normal pancreatic epithelial cells(P<0.05).The proliferation ability of PANC-1 and MIA PaCa-2 cells was significantly inhibited after over-expression of miR-1181(P<0.05),and the number of colony formation was significantly decreased(P<0.05).Subcutaneous tumor model in nude mice showed that the tumor size and immunohistochemistry index Ki67 were significantly lower than those in the normal control group(P<0.05) after over-expression of miR-1181.Conclusion:Over-expression of miR-1181 can inhibit the proliferation of pancreatic cancer cells in vitro and in vivo,and is expected to become a new target for the treatment of pancreatic cancer.

关键词(KeyWords)： 胰腺肿瘤;细胞;增殖;慢病毒感染;微小RNA;miR-1181
pancreatic neoplasms;cells;proliferation;lentivirus infections;micro RNA;miR-1181

Abstract:

Keywords:

基金项目(Foundation): 国家国际科技合作专项资助(2014DFA31420);; 国家自然科学基金(81160311,81572429,81660483)

作者(Author): 王杰;喻超;周显飞;朱昌豪;黄巍;江建新;
WANG Jie;YU Chao;ZHOU Xianfei;ZHU Changhao;HUANG Wei;JIANG Jianxin;Department of Hepatic-Biliary-Pancreatic Surgery,the Affiliated Hospital of Guizhou Medical University;

Email:

DOI: 10.19367/j.cnki.1000-2707.2016.12.004

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