皮层肌动蛋白的差异表达与结肠癌转移的相关性研究

目的 检测具有不同转移能力的结肠癌细胞亚系中皮层肌动蛋白的表达,探讨皮层肌动蛋白的差异表达与结肠癌转移的相关性.方法 本实验应用具有不同肝转移能力的结肠癌细胞系SW1116、人结肠癌皮下5代细胞系(SW1116_5)、人结肠癌肝转移细胞系1代(CRCLM1)和人结肠癌肝转移细胞系3代(CRCLM3).4种细胞在皮下成瘤后再原位种植到裸鼠结肠观察其肝脏转移能力的差异,用Western blotting和real time-PCR检测皮层肌动蛋白表达的差异,同时行体外迁移侵袭实验检测癌细胞的侵袭转移能力的差异,免疫组化方法分析临床86例结肠癌组织中皮层肌动蛋白的表达情况及其与临床病理特征之间的关系.结果 Western blotting和real time-PCR均证实在4种细胞中皮层肌动蛋白表达自SW1116、SW1116_5、CRCLM1、CRCLM3逐渐增高,四者两两比较差异有统计学意义,均P＜0.05;动物实验证实CRCLM3(100%)和CRCLM1(89%)与SW1116(40%)和SW1116_5(44%)相比具有较强的转移能力,免疫组化提示皮层肌动蛋白表达在86例中57例(66%)为强阳性,29例(34%)为阴性.结论 皮层肌动蛋白差异表达与结肠癌的转移以及临床分期呈正相关,皮层肌动蛋白有可能成为结肠癌临床分期以及肿瘤转移的判断指标.

The correlation between cortactin differential expression and metastasis of colorectal cancer

Objective To explore the correlation between cortactin differential expression and metastasis of colorectal cancer by testing the expression of cortactin in cell sublines with different metastatic ability. Method Human colon cancer cell line SW1116, and sublines of fifth subcutaneous ( SW1116_5), first, fifth generation (CRCLM1 、CRCLM3 ) characteristic of different liver metastasis were implanted subcutaneously in BALB/c mouse. Tumors were transplanted into the colon, and liver metastasis was observed, Western blotting and real time-PCR was used to detect the varience of cortactin. Transwell assay was applied to evaluate four cell lines migration and invasion ability. Immunohistochemistry was used to analyze the relationship of cortactin and clinicopathological characterizing in 86 cases of colorectal cancer.Result CRCLM3( 100％ )has higher ability of metastasis than that of CRCLM1 ( 89％ ), SW1116(40％ )and SW1116_5 (44％) respectively. Expression of cortactin in the SW1116, SW1116_5, CRCLM1,CRCLM3 was gradually increased. There were significant differences between four cell lines by comparison between each others ( P ＜ 0.05 ). Immunohistochemical expression of cortactin in 86 cases was positive in 57 cases(66％ ), and negative in 29 cases (34％). Conclusions Different expression of cortactin with colorectal cancer metastasis and clinical stage was positively correlated. Cortactin is a potential indicator for clinical staging and tumor metastasis of colorectal cancer.