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1504-siRNA对EphA3表达肿瘤细胞HCT116的抑制作用研究
引用本文:赵亚丽,武瑞琴,李海量,王洪涛,罗国兰,王健,周建光. 1504-siRNA对EphA3表达肿瘤细胞HCT116的抑制作用研究[J]. 军事医学, 2012, 36(4): 272-275
作者姓名:赵亚丽  武瑞琴  李海量  王洪涛  罗国兰  王健  周建光
作者单位:赵亚丽 (军事医学科学院生物工程研究所,北京,100850) ; 武瑞琴 (航天医学工程研究所,北京,100193) ; 李海量 (军事医学科学院疾病预防控制所,北京,100850) ; 王洪涛 (军事医学科学院生物工程研究所,北京,100850) ; 罗国兰 (军事医学科学院生物工程研究所,北京,100850) ; 王健 (军事医学科学院生物工程研究所,北京,100850) ; 周建光 (军事医学科学院生物工程研究所,北京,100850) ;
基金项目:国家自然科学基金资助项目
摘    要:
目的研究原癌基因EphA3的小干扰RNA(small interfering RNA)1504-siRNA对高表达结肠癌细胞HCT116的增殖抑制作用。方法将1504-siRNA质粒用Vigofect转染试剂瞬时转染HCT116细胞,36~48 h后,收集蛋白,用Western印迹检测EphA3及AKT信号通路中的蛋白分子表达,收集细胞,进行噻唑蓝(MTT)实验、平板克隆形成实验和软琼脂克隆形成实验。结果 1504-siRNA能抑制HCT116细胞的EphA3蛋白水平,抑制其增殖、平板克隆和软琼脂克隆的形成,抑制pmTOR、p-c-Raf、pAKT、p-4ebp1等信号分子的表达。结论 1504-siRNA抑制HCT116细胞的增殖、存活能力和恶性程度,这可能是通过抑制AKT信号通路来实现,它有望作为肿瘤治疗的候选药物。

关 键 词:EphA3  结肠肿瘤  增殖  RNA,小分子干扰  HCT116细胞

Inhibition effect of 1504-siRNA on EphA3 expressed HCT116 cells
ZHAO Ya-li,WU Rui-qin,LI Hai-liang,WANG Hong-tao,LUO Guo-lan,WANG Jian,ZHOU Jian-guang. Inhibition effect of 1504-siRNA on EphA3 expressed HCT116 cells[J]. Military Medical Sciences, 2012, 36(4): 272-275
Authors:ZHAO Ya-li  WU Rui-qin  LI Hai-liang  WANG Hong-tao  LUO Guo-lan  WANG Jian  ZHOU Jian-guang
Affiliation:1*(1.Beijing Institute of Biotechnology,Beijing 100850,China;2.Institute of Aerospace Medical Engineering,Beijing 100193,China;3.Institute of Disease Prevention and Control,Academy of Military Medical Sciences,Beijing 100850,China)
Abstract:
Objectives To investigate the inhibition effect of the small interfering RNA(siRNA) targeting EphA3,1504-siRNA,on proliferation of colon cancinoma cells HCT116.Methds 36-48 hours after plasmid carrying 1504-siRNA was transfected into HCT116 cells,protein was collected and the protein level of EphA3 and AKT signal molecules was tested by Western blotting.Furthermore,MTT,plate clonogenicity and soft agar clonogenicity experiment were accomplished.Results 1504-siRNA reduced the level of EphA3,pmTOR,p-c-Raf,pAKT and p-4ebp1,et al.It surpressed the ability of cell proliferation,plate clonogenicity and soft agar clonogenicity of HCT116 cells.Conclusion 1504-siRNA might be a promising drug for cancer gene therapy,for it can inhibit the proliferation,survival,and malignancy of EphA3 highly expressed tumor cells,which may result from the suppression of AKT signal pathway.
Keywords:EphA3  colonic neoplasms  proliferation  RNA,small interfering  HCT116 cells
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