| Hsa-miR-15a/16-1靶向抑制Bcl-2基因对鼻咽癌CNE-2细胞的抑制作用 |
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| 引用本文： | 张春鸿,方潇碧,林森,黄亚,施清圆,吴丽萍,黄振校,蒋磊,谭映霞,廖志苏. Hsa-miR-15a/16-1靶向抑制Bcl-2基因对鼻咽癌CNE-2细胞的抑制作用[J]. 中国耳鼻咽喉头颈外科, 2011, 0(10): 516-520 |
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| 作者姓名： | 张春鸿 方潇碧 林森 黄亚 施清圆 吴丽萍 黄振校 蒋磊 谭映霞 廖志苏 |
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| 作者单位： | 温州医学院附属第一医院耳鼻咽喉科;温州市第八人民医院耳鼻咽喉科;温州医学院附属第一医院医学科学研究所; |
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| 基金项目： | 浙江省科技计划项目(2010C34005); 浙江省温州市科技计划项目(Y20090044); 浙江省温州市鹿城区科技计划项目(S090103)联合资助 |
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| 摘 要： | 目的观察Hsa-miR-15a/16-1靶向抑制B细胞淋巴瘤/白血病-2基因（B-cell lymphoma gene 2,Bcl-2gene）表达对鼻咽癌（nasopharyngeal carcinoma,NPC）CNE-2细胞的生长抑制作用。方法将构建成功的重组质粒pENTR-CMV-EGFP-hsa-mir-15a/16-1经Lipofectamine 2000转染CNE-2细胞,RT-qPCR检测miR-15a/16-1、Bcl-2 mRNA表达水平,流式细胞术和WesternBlot检测Bcl-2、caspase-3蛋白表达水平,CCK-8法分析细胞生长抑制情况,4,6-二脒基-2-苯基吲哚（4,6-diamidino-2-phenylindole,DAPI）染色观察细胞凋亡状况。结果转染后miR-15a表达增高（F=547.525,P均〈0.001）;miR-16-1表达增高（F=99.979,P均〈0.001）;Bcl-2 mRNA表达无明显差别（F=0.220,P〉0.05）;Bcl-2蛋白表达下调,caspase-3蛋白表达增高;细胞在体外凋亡明显;细胞增殖受到抑制,作用呈明显的时效关系。结论 Hsa-miR-15a/16-1对Bcl-2基因的靶向抑制,可体外对CNE-2细胞产生增殖抑制和促进凋亡作用。
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| 关 键 词： | 鼻咽肿瘤 转染 细胞凋亡 流式细胞术 |
Inhibitory effect of nasopharyngeal carcinoma CNE-2 cells through targeted inhibitory effect on the Bcl-2 gene expression by Hsa-mir-15a/16-1 |
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| ZHANG Chunhong,FANG Xiaobi,LIN Sen,HUANG Ya,SHI Qingyuan,WU Liping,HUANG Zhenxiao,JIANG Lei,TAN Yingxia,LIAO Zhisu. Inhibitory effect of nasopharyngeal carcinoma CNE-2 cells through targeted inhibitory effect on the Bcl-2 gene expression by Hsa-mir-15a/16-1[J]. Chinese Archives of Otolaryngology-Head and Neck Surgery, 2011, 0(10): 516-520 |
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| Authors: | ZHANG Chunhong FANG Xiaobi LIN Sen HUANG Ya SHI Qingyuan WU Liping HUANG Zhenxiao JIANG Lei TAN Yingxia LIAO Zhisu |
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| Affiliation: | ZHANG Chunhong1,FANG Xiaobi1,LIN Sen1,HUANG Ya2,SHI Qingyuan1,WU Liping1,HUANG Zhenxiao1,JIANG Lei3,TAN Yingxia3,LIAO Zhisu1(1 Department of Otolaryngology,the First Affiliated Hospital of Wenzhou Medical College,Wenzhou,Zhejiang,325000,China,2 Department of Otolaryngology,the Eighth People's Hospital of Wenzhou,3 Medical and Scientific Institute of the First Affiliated Hospital of Wenzhou Medical College,China) |
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| Abstract: | OBJECTIVE To observe the inhibitory effect of Targeted inhibition of Bcl-2 gene expression by Hsa-miR-15a/16-1 on the growth of nasopharyngeal carcinoma CNE-2 cells.METHODS The recombinant plasmid pENTR-CMV-EGFP-hsa-mir-15a/16-1 which had been successfully constructed was transfected into CNE-2 cells with the help of Lipofectamine 2000.The expression of miR-15a/16-1,Bcl-2 mRNA was detected by RT-qPCR.The levels of Bcl-2 and Caspase-3 Protein were measured by Western blot and FCM.The inhibitory effect of Bcl... |
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| Keywords: | Nasopharyngeal Neoplasms Transfection Apoptosis Flow Cytometry |
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