短发夹状RNA对人肝癌细胞环氧合酶-2的抑制作用
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王华枫, 徐可树, 钱 伟, 汪步海
短发夹状RNA;COX-2;肝细胞癌,王华枫,徐可树,钱伟,汪步海,王华枫,通讯作者:,Silencingcyclooxygenase-2ge
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王华枫, 徐可树, 钱伟, 汪步海, 华中科技大学同济医学院附属协和医院消化内科 湖北省武汉市 430022
王华枫, 2006年华中科技大学同济医学院附属协和医院硕士, 主要从事肝癌的基础研究.
通讯作者: 徐可树, 430022, 湖北省武汉市, 华中科技大学同济医学院附属协和医院消化内科. xuzou@medmail.com.cn
电话: 027-67119890
收稿日期: 2006-07-03 接受日期: 2006-08-10
Silencing cyclooxygenase-2 gene by construction of eukaryotic expression vector expressing short hairpin RNA in human hepatocellular carcinoma cells
Hua-Feng Wang, Ke-Shu Xu, Wei Qian, Bu-Hai Wang
Hua-Feng Wang, Ke-Shu Xu, Wei Qian, Bu-Hai Wang,Department of Gastroenterology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China
Correspondence to:Dr. Ke-Shu Xu, Department of Gastroenterology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China. xuzou@medmail.com.cn
Received:2006-07-03 Accepted:2006-08-10
Abstract
AIM: To construct the eukaryotic expression plasmids of short hairpin RNA (shRNA) specific to human cyclooxygenase-2 (COX-2) gene and observe their inhibitory effects on COX-2 expression in human hepatocellular carcinoma cells.
METHODS: Plasmids named WBH1 and WBH2, containing the different sequences of human COX-2 mRNA coding region, were constructed. The expression of COX-2 was assayed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, respectively, 24, 48, 72 and 96 h after HepG2 and Bel7402 cells were transfected with liposomes.
RESULTS: The transfection rates in HepG2 and Bel7402 cells were about 60% and 54% respectively. RT-PCR showed that the inhibition efficiencies of the plasmid WBH1 were 18.5%, 88.6%, 52.8%, 42.4% in HepG2 cells and 9%, 45.1%, 70.1%, 56.3% in Bel7402 cells, respectively, 24, 48, 72 and 96 h after transfection (P < 0.01). Western blot demonstrated that the inhibition efficiencies of the plasmid WBH1 were 10 ......
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