Cortactin过表达对结肠癌细胞转移能力的影响

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)。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。

Effect of cortactin gene overexpression on metastases of colon cancer cells

Objective To evaluate the role of cortactin in metastases of human colon cancer cells.Methods The lentiviral vector with CTTN gene and green fluorescent protein was constructed and transfected into the 293T cells to produce mature lentivirus.The supernatant of virus-producing cells containing CTTN and GFP genes were then used to transfect SW1116 cells.Labeling protein GFP was detected by fluorescent microscopy.The expression of cortactin was detected by Western blot.Transwell assay was used to examine the change in the invasive ability of SW1116 cells.Furthermore,a mouse model of metastatic colon cancer was generated by following methods: SW1116 cells were transplanted into the subcutis of nude mice and then the subcutaneous tumor was implanted into the cecal wall of new nude mice to observe liver metastases.Results In the invasion and migration test,the number of cells crossing through chambers in the experimental group were(147±12) and(286±17),respectively,which were both higher than that in negative control group(83±10) and(112±11) and blank control group(75±7) and(103±9),P<0.05.The number of mice developing liver metastases in mice injected with recombinant lentiviral vector carrying CTTN gene transfected cells was significantly higher than that in mice injected with untransfected or empty plasmid transfected cells 7/12 vs 2/12 and 3/12,P<0.05).Conclusions Upregulating the expression of cortactin could significantly promote the invasion and migration of colon cancer cells.The result suggests that cortactin plays an important role in the metastases of colon cancer.