ENPP1诱导肿瘤干性和上皮间质转化促进口腔鳞癌的发展

ENPP1 promotes oral squamous cell carcinoma progression by inducing epithelial-mesenchymal transition phenotypes and stem cell features

目的癌症干细胞(cancer stem cell,CSC)特征和上皮间质转化(epithelial-mesenchymal transition,EMT)在口腔鳞癌(oral squamous cell carcinoma,OSCC)的进展和转移中至关重要。本研究旨在探讨ENPP1在调控口腔鳞癌细胞干性和转移中的重要作用。方法采用免疫组织化学染色和免疫印迹检测口腔鳞癌肿瘤组织及正常癌旁组织中ENPP1的表达及分布。通过免疫印迹检测口腔鳞癌细胞系(SCC-9,CAL-27和Tca-8113)和正常人口腔粘膜细胞系中ENPP1的表达。建立CAL-27 ENPP1敲低细胞系(shENPP1-#1和shENPP1-#2),通过免疫印迹检测CSC标志物,并通过划痕实验和免疫荧光实验检测肿瘤细胞迁移能力及EMT转化。结果通过临床样本检测发现多数口腔鳞癌患者肿瘤组织中ENPP1的表达显著上调。细胞实验发现ENPP1在口腔鳞癌CAL-27和Tca-8113细胞中表达上调。CAL-27-shENPP1敲低细胞系中CSC标志物的表达显著降低,包括ABCG2,SOX2,NANOG和CD44。此外,ENPP1基因敲低逆转了TGFβ诱导的EMT及细胞迁移。结论ENPP1在诱导和维持口腔鳞癌细胞的CSC特征和EMT中起重要作用,可能为口腔鳞癌的临床治疗提供新靶点。

Objective Cancer stem cell(CSC)characteristics and epithelial-mesenchymal transition(EMT)are crucial in the progression and metastasis of oral squamous cell carcinoma(OSCC).The purpose of this study was to investigate the important role of ENPP1 in the regulation of the stem cell features and metastasis of OSCC.Methods Immunohistochemical staining and western blot were used to detect the expression and distribution of ENPP1 in tumor tissues of OSCC patients.The expression of ENPP1 in OSCC cell lines(SCC-9,CAL-27 and Tca-8113)and normal human oral mucosal cell lines was detected and establish CAL-27 ENPP1 knockdown cell lines(shENPP1-#1 and shENPP1-#2).CSC markers were detected by immunoblotting,and tumor cell migration ability and EMT transformation were detected by wound healing assay and immunofluorescence.Results The expression of ENPP1 in tumor tissue of most patients with OSCC was significantly up-regulated.In vitro study showed that ENPP1 was up-regulated in CAL-27 and Tca-8113 cells lines.The expression of CSC markers in CAL-27-shENPP1 knockdown cell lines was significantly reduced,including ABCG2,SOX2,NANOG,and CD44.In addition,ENPP1 gene knockdown reversed TGFβ-induced EMT and cell migration.Conclusion ENPP1 plays an important role in inducing and maintaining the stem cell features and EMT of oral squamous cell carcinoma,and may provide a new target for clinical treatment.