结直肠癌组织中NEK2的表达及其对HCT116细胞增殖、侵袭和迁移能力的影响

Expression of NEK2 in Colorectal Cancer Tissue and Its Effect on Proliferation,Invasion and Migration of HCT116 Cells

目的探讨中心体相关激酶2(NEK2)在结直肠癌组织中的表达及对结直肠癌细胞HCT116增殖、侵袭及迁移的影响。方法构建针对NEK2的小干扰RNA(si-NEK2),脂质体法转染HCT116细胞。CCK8实验、流式细胞术、划痕实验和Transwell小室实验检测敲低NEK2表达后细胞增殖、周期分布、侵袭和迁移能力的改变。Western blot检测敲低NEK2表达后相关蛋白表达水平的改变。结果NEK2在结直肠癌组织中的阳性表达率明显高于癌旁组织(65.0%vs.35.0%,χ~2=14.593,P<0.01),其表达水平与结直肠癌TNM分期、淋巴结转移和远处转移显著相关(均P<0.05);NEK2蛋白及mRNA在结直肠癌细胞中的表达水平显著高于正常结直肠黏膜细胞(P<0.01)。敲低NEK2表达后,HCT116细胞出现明显的G_0/G_1期阻滞,细胞的增殖、侵袭及迁移能力均显著降低(P<0.01),E-cadherin表达含量升高,N-cadherin、CDK4和cyclin D1表达含量降低。结论NEK2在结直肠癌组织中高表达并与临床病理特征相关,下调NEK2表达可有效抑制人结直肠癌细胞的增殖、侵袭和迁移能力。提示NEK2在结直肠癌的发生发展过程中发挥着重要的作用,可作为潜在的治疗靶点。

Objective To investigate NIMA-related kinase 2(NEK2)expression in colorectal cancer(CRC)tissues and its effect on the proliferation,migration and invasion of HCT116 cells.Methods Synthesized siRNA targeting NEK2 was transfected into HCT116 cells by lipofectamine?method.CCK8 assay,FACS,wound healing assay and Transwell assay were used to detect the effects of NEK2 knockdown on cell proliferation,cell cycle distribution,migration and invasion.Western blot was carried out to detect the expression of E-cadherin,N-cadherin,CDK4 and cyclin D1.Results The expression of NEK2 in CRC tissues was significantly higher than that in adjacent normal tissues(65.0%vs.35.0%,χ2=14.593,P<0.01),and its level was closely related to TNM stage,lymph node metastasis and distant metastasis(all P<0.05).NEK2 protein and mRNA levels in CRC cell lines were also significantly higher than those in normal colorectal mucosal cells(P<0.01).After NEK2 was knocked down by siRNA,HCT116 cells were arrested in G0/G1 cycle,while cell proliferation,invasion and migration were significantly reduced(P<0.01).NEK2 interference in HCT-116 cells led to the upregulation of E-cadherin and downregulation of N-cadherin,CDK4 and cyclin D1 at protein levels.Conclusion The high NEK2 expression is correlated with clinicopathological characteristics of CRC patients.NEK2 knockdown by siRNA could effectively inhibit the proliferation,invasion and migration abilities of colorectal cancer cells,suggesting that NEK2 plays an important role in the occurrence and development of colorectal cancer and it can be used as a potential treatment target.