滋养层细胞表面抗原-2促进鼻咽癌细胞增殖、迁移和侵袭能力的实验研究

Effects of trophoblast cell-surface antigens 2 on nasopharyngeal carcinoma proliferation,migration and invasion

目的研究滋养层细胞表面抗原-2(Trop2)对鼻咽癌CNE-2细胞增殖、迁移和侵袭能力的影响及可能的调控机制。方法先应用免疫组织化学方法检测Trop2在鼻咽癌组织及对照鼻咽上皮中的表达。随后在鼻咽癌CNE-2细胞中,应用转染技术分别上调和下调Trop2的表达,通过EdU染色、四甲基偶氮唑蓝(MTT)法、实时荧光定量聚合酶链反应、细胞迁移实验(Transwell法)、细胞侵袭实验检测Trop2对鼻咽癌细胞行为学的影响,并采用Western印迹技术检测Trop2对E-cadherin、N-cadherin蛋白表达的影响。结果Trop2在鼻咽癌组织中高表达,且与患者的淋巴结转移及肿瘤分期显著相关。Trop2过表达可以上调鼻咽癌细胞的增殖能力和鼻咽癌细胞中干细胞基因Nanog、OCT4 mRNA的表达。Trop2过表达可以增强鼻咽癌细胞的迁移和侵袭能力。与对照组相比,高表达Trop2细胞组E-cadherin表达下调,N-cadherin表达上调,低表达Trop2细胞组则相反。结论Trop2在鼻咽癌组织中表达增加,可能是鼻咽癌肿瘤干细胞的特征基因,其可能通过诱导上皮质细胞-间充质转化过程促进鼻咽癌细胞的迁移和侵袭。

Objective To investigate the roles of trophoblast cell-surface antigens-2(Trop2)in the proliferation,migration and invasion of nasopharyngeal carcinoma.Methods The expressions of Trop2 in nasopharyngeal carcinoma and counterparts in normal nasopharyngeal tissue were investigated by immunohistochemistry.Trop2 was knocked down or over-expressed by transfected with recombinant adenoviral in CNE-2 cell line of nasopharyngeal carcinoma.Methyl thiazolyl tetrazolium(MTT),EdU labeling,real-time quantitative polymerase chain reaction(RT-PCR)and Transwell test were used to explore the roles of Trop2 in praxiology.The expression levels of Trop2,E-cadherin and N-cadherin protein in CNE-2 cells after transfection were determined by Western blot assays.Results The expressions of Trop2 were significantly upregulated in nasopharyngeal carcinoma tissues and correlated with tumor classification and lymph node metastasis.Overexpression of Trop2 significantly upregulated the cell proliferation and expressions of Nanog,OCT4 mRNA in CNE-2 cells.Transwell assay showed that Trop2 promoted the migration and invasion abilities of CNE-2 cells.Furthermore,Western blot assay showed that overexpression of Trop2 significantly increased the expressions of N-cadherin,reduced the expressions of E-cadherin,the effect of Trop2 knock-down exerted the opposite effect.Conclusions Trop2 was significantly upregulated in nasopharyngeal carcinoma tissues,which might play an important role in nasopharyngeal carcinoma cancer stem cell and promoted EMT-induced nasopharyngeal carcinoma migration and invasion.