趋化性细胞因子受体CXCR4基因沉默对大肠癌细胞体外侵袭及增殖能力的影响
CXCR4基因;短干扰RNA;结直肠癌;侵袭;逆转录-聚合酶链反应;免疫印迹技术;Boyden小室模型;流式细胞术;MTT法,张孟贤,韩娜,冷彦,张孟贤,通讯作者:,Effectofspecificsuppressio
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张孟贤, 韩娜, 冷彦, 华中科技大学同济医学院附属同济医院肿瘤科 湖北省武汉市 430030
张孟贤, 2001年华中科技大学同济医学院硕士研究生毕业, 2005年获博士学位, 主治医师, 主要从事恶性肿瘤的侵袭和转移研究.
国家自然科学基金资助项目, No. 30570961
通讯作者: 冷彦, 430030, 湖北省武汉市解放大道1095号, 华中科技大学同济医学院附属同济医院肿瘤科. yan_leng@126.com
电话: 027-83663360
收稿日期: 2007-01-28 接受日期: 2007-02-09
Effect of specific suppression of chemokine receptor CXCR4 by short interference RNA on the invasion capability and proliferation of colorectal cancer cells
Meng-Xian Zhang, Na Han, Yan Leng
Meng-Xian Zhang, Na Han, Yan Leng,Department of Oncology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China
Supported byNational Natural Science Foundation of China, No. 30570961
Correspondence to:Yan Leng, Department of Oncology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, 1095 Jiefang Road, Wuhan 430030, Hubei Province, China. yan_leng@126.com
Received:2007-01-28 Accepted:2007-02-09
Abstract
AIM: To study the inhibitory effect of chemokine receptor CXCR4 short interference RNA (siRNA) on the invasion capability and proliferation of colorectal cancer cell line SW480.
METHODS: SiRNA specifically targeting CXC chemokine receptor CXCR4 was designed in vitro using T7 RNA polymerase. Colorectal cancer SW480 cells were cultured under standard condition and the siRNA was transfected into SW480 cells with Lipofectamine 2000. Negative control and mock control were used at the same time. The levels of CXCR4 mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting 48 h after transfection respectively. The changes of invasion capability and MT1-MMP protein in SW480 cells were evaluated using Boyden Chamber invasion assay and Western blotting respectively. The cell cycle distribution and proliferation status were detected by flow cytometry and MTT assay respectively ......
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