摘要
目的研究结肠癌组织CCAAT/增强子结合蛋白-α(C/EBPα)的表达,及其过表达对结肠癌SW480细胞系侵袭性的影响。方法利用免疫组化方法检测48例结肠癌患者的癌组织与正常组织C/EBPα表达情况,并统计分析C/EBPα的表达与结肠癌临床病理参数之间的关系。构建p CDGFP-C/EBPα真核表达质粒,通过细胞划痕实验观察转染对结肠癌SW480细胞系迁移能力的影响,并检测与肿瘤侵袭相关蛋白KLF5、MMP-2、MMP-9、ECD的的表达。结果免疫组化结果显示,正常组织C/EBPα的低表达率为6.25%,而结肠癌组织C/EBPα的低表达率为68.75%,两组比较差异有统计学意义(P<0.05),且低表达C/EBPα的患者肿瘤直径越大、TMN分期越晚、淋巴结转移概率越大。细胞划痕实验显示,划痕48h、72h后,正常SW480细胞的迁移率分别为53.16%、80.77%,而过表达C/EBPα的SW480细胞的迁移率分别为23.57%、31.89%,两组比较差异有统计学意义(P<0.05);免疫印记实验表明,过表达C/EBPα的SW480细胞的KLF5、MMP-2、MMP-9表达明显降低,而ECD表达明显升高。结论 C/EBPα在结肠癌组织低表达,且与结肠癌的TMN分期和转移有密切关系,C/EBPα过表达能够显著降低结肠癌细胞的侵袭性,对今后结肠癌的早期诊断和基因靶向治疗有重要指导意义。
Objective To study the expression of C/EBPα in colon cancer tissues and the effect of its over-expression on invasion of SW480. Methods The expression level of C/EBPα was detected in cancer tissues and normal tissues from 48 patients with colon cancer by immunohistochemistry. The relationships between theexpression of C/EBPα and clinicopathological parameters of colon cancer were statistically analyzed. p CDGFP-C/EBPα eukaryotic expression vector was constructed and transfected into SW480 to study the change of invasive ability of tumor cells by wound scratch assay and to detect the change of tumor invasion-associated protein（KLF5, MMP-2, MMP- 9,ECD） expressions. Results Immunohistochemistry results showed that the rate of low expression of C/EBPα was6.25% in normal tissues and 68.75% in colon cancer tissues, with a statistically significant difference between them（P〈0.05）. Patients with low expression of C/EBPα hada larger tumor diameter, later stages of the TMN and higherprobability of lymph node metastasis. Wound scratch assay showed that the migration rate of normal SW480 cells was 53.16% after 48 h and 80.77% after 72h（P 0.05）; the migration rate of SW480 cells that overexpressed C/EBPα was 23.57%after 48 h and 31.89% after 72h（P〈0.05）. Immunoblotting experiments also showed that the expression of KLF5, MMP-2, MMP-9 was significantly decreased, but ECD was significantly increased in SW480 cells with overexpressed C/EBPα. Conclusion The expression of C/EBPα reduces in colon cancer, and it has a close relationship with TMN staging and migration. The overexpression of C/EBPα can significantly reduce the invasion of colon cancer cells and hence it may have a great significance on early diagnosis and targeted therapy of colon cancer in the future.
出处
《浙江中西医结合杂志》
2016年第2期113-116,共4页
Zhejiang Journal of Integrated Traditional Chinese and Western Medicine
基金
浙江省乐清市软课题与社会发展项目(No.2014Y005)
