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目的:通过诱导皮层肌动蛋白(cortactin)的过表达分析其在结肠癌细胞侵袭迁移以及肝转移过程中的作用。方法:构建含CTTN基因和绿色荧光蛋白的慢病毒表达载体,转染293T细胞。取含CTTN基因和绿色荧光蛋白的病毒上清液转染结肠癌细胞株SW1116使cortactin稳定表达。蛋白印迹实验检测cortactin的表达,通过Transwell实验评估结肠癌细胞侵袭迁移能力的改变。利用SW1116细胞株使裸鼠皮下成瘤,取肿瘤组织种植新一批裸鼠结肠壁并观察结肠癌肝转移的情况。结果:成功构建cortactin过表达的SW1116细胞。侵袭和迁移实验中,过表达组穿膜细胞数分别为(147±12)个、(286±17)个,而阴性对照组为(83±10)个、(112±11)个,空白对照组为(75±7)个、(103±9)个,后两组与过表达组相比差异均有统计学意义(P<0.05)。动物实验显示cortactin过表达组肝转移能力明显高于阴性对照组和空白对照组(7/12比2/12和3/12)(P<0.05)。结论:上调cortactin的表达能显著促进结肠癌细胞的侵袭迁移和肝转移能力,提示cortactin在结肠癌的转移过程中发挥重要作用。
OBJECTIVE: To investigate the role of cortactin in invasion and migration of colon cancer cells and liver metastasis by inducing cortactin overexpression. Methods: Lentivirus expression vector containing CTTN gene and green fluorescent protein was constructed and transfected into 293T cells. Cortactin was stably expressed by transfecting colon cancer cell line SW1116 with virus supernatant containing CTTN gene and green fluorescent protein. The expression of cortactin was detected by Western blotting and the invasion and migration of colon cancer cells were evaluated by Transwell assay. Subcutaneous tumor formation in nude mice was induced by SW1116 cell line, and a new group of nude mouse colon walls were planted and tumor metastasis of colon cancer was observed. Results: SW1116 cells overexpressing cortactin were successfully constructed. In the invasion and migration experiments, the numbers of transmembrane cells in the over-expression group were (147 ± 12) and (286 ± 17), respectively, while those in the negative control group were (83 ± 10) and (112 ± 11) (75 ± 7) and (103 ± 9), respectively. There was significant difference between the latter two groups and the overexpression group (P <0.05). Animal experiments showed that liver metastasis of cortactin overexpression group was significantly higher than that of negative control group and blank control group (7/12 vs 2/12 and 3/12) (P <0.05). CONCLUSION: Upregulation of cortactin can significantly promote invasion and migration of colon cancer cells and liver metastasis, suggesting that cortactin may play an important role in the metastasis of colon cancer.