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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 small cell interfering RNA (siRNA) of cell divisioncy cle42 (Cdc42) on the malignant phenotype of human colon cancer cells.Methods Western blot was used to detect the expression of Cdc42 in five colon cancer cell lines. Three pairs of siRNAs targeted to Cdc42 coding region and negative control RNA were designed and synthesized. LipofectamineTM2000 was transfected into Lovo cells and SW620 cells with high expression of Cdc42 in colon cancer cell lines. The expression of Cdc42 mRNA and protein were detected by RT-PCR and Western- expression. Cell Counting Kit-8 was used to detect the cell proliferation, scratch and scratching experiments and Transwell chamber assay were used to detect cell migration and invasion ability changes. Results The results of RT-PCR and Western blot showed that Cdc42-siRNA significantly down-regulated Cdc42mR-NA and protein levels, especially Cdc42-siRNA1. After siRNA treatment, the proliferation of tumor cells was inhibited and the ability of cell migration and invasion was significantly decreased. Conclusion Cdc42-siRNA can effectively inhibit the proliferation, migration and invasion of colon cancer cells. These results suggest that Cdc42 may be a new molecular target of inhibiting colon cancer cell proliferation and metastasis.