靶向cripto siRNA转染对裸鼠实验性大肠癌细胞肝转移的影响

Effects of cripto siRNA on liver metastasis of colorectal cancer cell in nude mice

目的研究cripto基因对人大肠癌细胞迁移、侵袭和肝转移的影响。方法以cripto基因mRNA小干扰RNA(small interfering RNA,siRNA)转染人大肠癌SW480细胞后,分别采用荧光实时定量RT-PCR和Western blot检测cripto基因mRNA和蛋白水平,以划痕试验方法评测癌细胞迁移能力;用Boyden模型观察癌细胞侵袭能力。以脾切除法建立裸鼠大肠癌肝转移模型,以cripto siRNA转染48h后的癌细胞接种裸鼠,观察对大肠癌细胞体内肝转移的影响。结果siRNA转染组cripto基因mRNA水平明显降低,且呈浓度和时间依赖性(P<0.001;P<0.001)。体外实验发现,cripto siRNA转染组癌细胞迁移距离和穿膜细胞数目明显低于对照组,且呈浓度依赖性(P<0.05)。体内实验发现,cripto基因siRNA转染组SW480细胞转移率和肿瘤结节数均明显低于对照组(P<0.05;P<0.05)。结论cripto基因在大肠癌肝转移中起着重要的作用;以siRNA下调cripto基因表达,可抑制大肠癌细胞肝转移。

Objective To study the effects of cripto on migration, invasion, and liver metastasis of colorectal cancer cell. Methods After human colorectal cancer cell line SW480 was transfected by cripto small interfering RNA （siRNA）, the mRNA and protein level were determined by Real-time RT-PCR and Western blot, respectively. The migration and invasion ability were evaluated by wound-healing assay and boyden chamber model, respectively. Thirty nude mice model of liver metastasis from colorectal cancer was established by splenectomy. Results The siRNA could down-regulate the level of mRNA and protein of cripto in a dose- and time-dependent manner. Suppression of cripto expression could inhibit migration and invasion ability of human colorectal cancer cell in vitro. The metastastic rate and tumor nodules were lower in transfection with cripto siRNA than in two control groups in vivo. Conclusions Cripto gene might play an important role in regulation of liver metastasis from colorectal carcinoma cell, and suppression of cripto gene by siRNA can inhibit liver metastasis of colorectal cancer.