CDC20、TOP2A、NEK2的表达特征以及对食管鳞状细胞癌细胞增殖和迁移的影响

Expression characteristics of CDC20,TOP2A and NEK2 and their effects on proliferation and migration of esophageal squamous cell carcinoma cells

目的分析食管鳞状细胞癌(ESCC)中CDC20、TOP2A、NEK2基因表达与分期和预后的关系,讨论3者对ESCC细胞的增殖和迁移能力的影响,旨在寻找理想的生物标志物和治疗靶点。方法收集2011年1月至2017年12月鄂州市中心医院肿瘤科112例ESCC患者的临床资料和组织。利用实时荧光定量PCR、蛋白免疫印迹和免疫组织化学法分析CDC20、TOP2A、NEK2基因的表达差异;通过随访对患者进行生存预后分析;培养ESCC细胞系,对细胞分别转染过表达CDC20、TOP2A、NEK2的重组载体pcCDC20、pcTOP2A、pcNEK2,CCK-8实验和伤口愈合实验分析ESCC的增殖率和迁移率变化。结果与癌旁非癌组织比,ESCC组中CDC20、TOP2A、NEK2的mRNA和蛋白水平均增高,差异均有统计学意义(均P<0.05)。CDC20、TOP2A的高表达与淋巴结转移和生存预后差相关(均P<0.05);NEK2的高表达与肿瘤组织大小相关(P<0.05)。最后,分别与各组阴性对照组比,pcCDC20、pcTOP2A、pcNEK2组的细胞增殖率和迁移率均升高,差异均有统计学意义(均P<0.05)。结论CDC20、TOP2A、NEK2在ESCC组织中表达上调,3者均与ESCC的预后生存差相关;CDC20、TOP2A、NEK2均促进ESCC细胞的增殖和迁移。CDC20、TOP2A、NEK2基因均可能是ESCC的潜在诊断和预后生物标志物。

Objective To systematically analyze the relationship between CDC20,TOP2A,and NEK2 gene expression and stage and prognosis in esophageal squamous cancer(ESCC),discuss the effect of the three genes on the proliferation and migration of ESCC cells,and seek to find ideal biomarkers and therapeutic targets.Methods The clinical data and organization of 112 patients with ESCC were collected.The clinical data and tissues of 112 patients with ESCC in the Department of Oncology,Ezhou Central Hospital from January 2011 to December 2017 were collected.Real-time fluorescent quantitative PCR,Western blotting and immunohistochemistry methods were used to analyze the expression of CDC20,TOP2A,NEK2 genes.ESCC patients were followed up to analyze the survival prognosis.The ESCC cell line was cultured.The recombinant vectors pcCDC20,pcTOP2A,and pcNEK2 were transfected into the ESCC cells to overexpress CDC20,TOP2A,and NEK2,respectively.The CCK-8 experiment and wound healing experiment were used to analyze the changes in the proliferation rate and migration rate of ESCC cells.Results Compared with non-cancerous tissues adjacent to cancer,the mRNA and protein levels of CDC20,TOP2A,and NEK2 in the ESCC group increased,the differences were statistically significant(P<0.05).The high expression of CDC20 and TOP2A was associated with lymph node metastasis and poor survival prognosis(P<0.05).The high expression of NEK2 is related to the size of tumor tissue(P<0.05).Finally,compared with the negative control group of each group,the cell proliferation rate and migration rate of the pcCDC20,pcTOP2A,and pcNEK2 groups increased,the differences were statistically significant(P<0.05).Conclusion CDC20,TOP2A and NEK2 are up-regulated in ESCC tissues,and all three are associated with poor prognostic survival of ESCC;CDC20,TOP2A,NEK2 all promote the proliferation and migration of ESCC cells.CDC20,TOP2A and NEK2 genes may be potential biomarkers for diagnosis and prognosis of ESCC.