Cdc42小干扰RNA对结肠癌细胞恶性表型的影响  

Effects of Cdc42-siRNA on Malignant Phenotypes of Colon Cancer Cells

作　　者：刘素蕊[1,2] 张思源[1,2] 刘明[1] 朱亚杰[2] 周继陶[2] 黄娟[1,2] 唐秋琳[1] 陈向征[1] 郎楠[1] 毕锋[1,2] 

机构地区：[1]四川大学生物治疗国家重点实验室/信号转导及分子靶向治疗研究室,成都610041 [2]华西医院腹部肿瘤科

出　　处：《肿瘤防治研究》2009年第12期1016-1019,共4页Cancer Research on Prevention and Treatment

基　　金：国家自然科学基金资助项目(30672378);教育部博士点基金资助项目(20060610068)

摘　　要：目的研究细胞周期分裂蛋白Cell divisioncy cle42(Cdc42)小干扰RNA(siRNA)对人结肠癌细胞恶性表型的影响.方法Western blot检测五种结肠癌细胞系中Cdc42的表达。设计并合成靶向Cdc42编码区的三对siRNA及阴性对照RNA,应用LipofectamineTM2000分别转染结肠癌细胞系中高表达Cdc42的Lovo和SW620细胞,RT-PCR和Western-blot分别检测48h后Cdc42mRNA及蛋白的表达。Cell Counting Kit-8检测细胞的增殖能力,损伤刮擦实验和Transwell小室法分别检测细胞迁移与侵袭能力的变化。结果RT-PCR和Western blot检测显示,Cdc42-siRNA能显著下调Cdc42mR-NA和蛋白水平,尤其Cdc42-siRNA1;siRNA处理后的肿瘤细胞增殖受到抑制,细胞迁移、侵袭能力也显著降低。结论Cdc42-siRNA可有效抑制结肠癌细胞的增殖、迁移和侵袭。提示Cdc42可能成为抑制结肠癌细胞增殖和转移新的分子靶点。Objective To study the effect of cell division cycle 42（Cdc42） small interfering RNA（siRNA） on the malignant phenotype of human colon cancer cells.Methods The protein expression levels of Cdc42 in five colon cancer cell lines were examined with Western blot.Three Cdc42 sequence-specific small interfering RNA and negative control RNA were synthesized and transfected into Lovo and SW620 cells which have high expression of Cdc42 by LipofectamineTM2000.The expression of Cdc42 mRNA and protein were examined with RT-PCR and Western blot respectively after 48h. Cell growth rate was meas- ured by Cell Counting Kit-8. Wound healing and invasion assays were used to examine the abilities of mi gration and invasion of the cells, respectively. Results Both RT-PCR and Western blot revealed that Cdc42-siRNA notably dowwregulated Cdc42 expression at mRNA levels, especially the Cdc42 siRNA1. Western blot also revealed notably down-regulated Cdc42 expression at protein levels. The proliferation of colon cells was inhibited after iRNA treatment. The migrating and invasive abilities of the cells were also suppressed. Conclusion Cdc42 siRNA could effectively suppress the proliferation, migration and invasion of colon cancer cells. Cdc42 might be a potential target for molecular targeting therapy.

关 键 词：结肠癌 CDC42 RHO SIRNA 

分 类 号：R735.35[医药卫生—肿瘤] R73-35[医药卫生—临床医学]