摘要
目的:探讨RNA干扰技术沉默Annexin Ⅱ基因表达对人乳腺癌细胞MDA-MB-231侵袭迁移力的影响。方法:体外化学合成Annexin Ⅱ基因序列特异性双链小干扰RNA(Annexin Ⅱ-siRNA),转染高转移人乳腺癌细胞株MDA-MB-231,用荧光显微镜观察转染效率,逆转录-聚合酶链反应和蛋白印迹检测Annexin Ⅱ mRNA和蛋白表达水平;通过生长曲线和体外侵袭迁移实验,观察乳腺癌细胞恶性生物学行为的改变。结果:Annexin Ⅱ-siRNA能有效抑制Annexin Ⅱ mRNA和蛋白表达(P<0.05);经siRNA处理的细胞,细胞生长速度明显降低(P<0.05),侵袭迁移力显著下降(P<0.05)。结论:沉默Annexin Ⅱ基因表达可抑制乳腺癌细胞MDA-MB-231的侵袭迁移力。
Objectives：To investigate the effects of small interference RNA（siRNA） silencing Annexin Ⅱ on invasion and metastasis of human breast cancer cell MDA-MB-231.Methods：The chemically-synthetic siRNA duplexes targeting Annexin Ⅱ（Annexin Ⅱ-siRNA） was transiently transfected into MDA-MB-231 cells which have high metastatic potential by lipofectamin 2000.The transfection efficiency was observed by fluorescence microscopy.RT-PCR and western blot were used to semi-quantify the Annexin Ⅱ mRNA and protein levels.The malignant characters of transfected MDA-MB-231 cells including celluar proliferation rate,the activities of invasion and metastasis were analyzed by use of 3-（4,5-dimethylthiazol-2-yl）-5-diphenyl tetrazolium bromide（MTT） and millicell chamber assay,respectivilly.Results：Annexin Ⅱ-siRNA effectively inhibited Annexin Ⅱ mRNA and protein levels（P〈0.05）.With siRNA interferring cells,the ability of cells proliferation was decreased（P〈0.05）,and cells invasive and metastasis were depressed（P〈0.05）.Conclusion：Silencing Annexin Ⅱ could significantly inhibit the abilities of invasion and metastasis of human breast cancer cells MDA-MB-231 in vitro
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2009年第8期984-987,共4页
Journal of Chongqing Medical University
基金
重庆市自然科学基金(2006BB5427)
