VEGFR-3siRNA对人结肠癌细胞生长和侵袭力的影响  

INHIBITORY EFFECT OF VEGFR-3 siRNA ON GROWTH AND INVASION OF HUMAN COLON CANCER CELLS

作　　者：贾如江[1] 侯丽艳[2] 郝秋芳[3] 刘奇[1] 冯运章[1] 张有成[4] 

机构地区：[1]河北省邯郸市中心医院普外二科,河北邯郸056002 [2]河北省邯郸市中心医院ICU室,河北邯郸056002 [3]河北省邯郸市中心医院核医学科,河北邯郸056002 [4]兰州大学第二医院普外科,甘肃兰州730003

出　　处：《河北医科大学学报》2009年第7期680-684,共5页Journal of Hebei Medical University

摘　　要：目的探讨不同浓度血管内皮生长因子受体-3(vascular endothelial growth factor receptor-3,VEGFR-3)小分子干扰RNA(small interfering RNA,siRNA)对人结肠癌细胞生长和侵袭力的影响。方法设计合成VEGFR-3siRNA,以不同剂量转染到人结肠癌细胞株(human colon adenocarcinoma cell line,LoVo)细胞中,采用四甲基偶氮唑蓝显色法[3-(4,5 dimethl-2 thiahiazoyi)-3,5-di-phenyl-tetrazolium bromide,MTT]观察细胞生长变化,应用半定量逆转录聚合酶链反应、Western-blot检测转染前后VEGFR-3 mRNA和VEGFR-3蛋白表达的变化,基质黏附实验检测细胞转染后黏附能力,体外侵袭试验检测转染后肿瘤细胞侵袭性的改变。结果 VEGFR-3 siRNA引起LoVo细胞生长抑制,在一定范围内呈浓度、时间依赖性;转染siRNA后VEGFR-3 mRNA和VEGFR-3蛋白表达下降;转染siRNA 72 h后LoVo细胞的黏附能力显著下降,与对照组相比差异均有统计学意义(P<0.05);VEGFR-3 siRNA使LoVo细胞侵袭能力明显下降,LoVo细胞穿膜细胞数明显低于空白对照组、非特异性对照组(P <0.05)。结论 VEGFR-3siRNA能够在LoVo细胞中引发RNA干扰效应,下调VEGFR-3基因的表达可以抑制细胞增殖,使LoVo细胞黏附力和侵袭能力明显下降。Objective To investigate the effect of the vascular endothelial growth factor receptor-3(VEGFR-3)small interfering RNA(siRNA)in down-regulating the expression of VEGFR-3 and inhibition of cell growth and cell invasion in human colon cancer cell line,human colon adenocarcinoma cell line(LoVo)cell.Methods Designing and synthesising one pair of 21 bp siRNA targeting VEGFR-3 mRNA;LoVo cells were transfected with VEGFR-3 siRNA in different dosage;MTT were used to measure cell growth;The expressions of VEGFR-3 mRNA and protein were detected by semi-quantitative real time reverse transcriptase polymerase chain reaction(RT-PCR)and Western blot methods,respectively;The tumor adhesion ability was detected by cell-matrix adhesion experiment,and invasion ability was evaluated by Boyden chamber model.Results VEGFR-3 siRNA can inhibit the proliferation of human colon cancer cell line LoVo cells.In a certain range,the higher the concentration and the longer the acting time of VEGFR-3 siRNA,the stronger the inhibitory effects on cancer cells growth.After siRNA transfecting into LoVo cells,the expression of VEGFR-3 mRNA and protein was significantly inhibited.The VEGFR-3 siRNA reduced the adhesion ability of LoVo cells in comparison with the untransduced group and negative control group.The invasion assay demonstrated that the number of LoVo cells in transfection group was signifcantly lower than that in untransduced group and negative control group.Conclusion The VEGFR-3siRNA can inhibit cells proliferation;The siRNA of VEGFR-3 gene can effectively inhibit the mRNA and protein expression of VEGFR-3. The VEGFR-3siRNA can restrain the adhesion and invasion ability of LoVo cells.

关 键 词：结肠肿瘤 RNA干扰 生长 

分 类 号：R735.35[医药卫生—肿瘤]