重组人源TNC诱导胰腺癌细胞系PANC1的EMT及迁移和侵袭  

Recombinant human TNC induces EMT,migration and invasion of pancreatic cancer cell line PANC1

作　　者：蔡隽 沈文远 王慧[2] 谭珵 赵秀梅[1] 沈啸洪[2] CAI Jun;SHEN Wen-yuan;WANG Hui;TAN Cheng;ZHAO Xiu-mei;SHEN Xiao-hong(Centre for Research and Development of Anti Tumor Drugs,Tianjin Institute of Medical and Pharmaceutical Science,Tianjin 300020;School of Medicine,Nankai University,Tianjin 300071;Department of Orthopaedics,Tianjin Medical University General Hospital,Tianjin 300052,China)

机构地区：[1]天津市医药科学研究所肿瘤药物研发中心,天津300020 [2]南开大学医学院,天津300071 [3]天津医科大学总医院骨科,天津300052

出　　处：《基础医学与临床》2020年第2期167-172,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(31370861)

摘　　要：目的构建人肌腱蛋白C(TNC)真核表达质粒,利用稳定过表达TNC的胰腺癌细胞系以及探索TNC在胰腺癌转移中的作用及分子机制。方法采用独特的引物设计方法构建TNC表达质粒,经转染和G418筛选获得稳定过表达TNC的胰腺癌PANC1细胞系。通过RT-qPCR和Western blot对细胞系中TNC的表达进行鉴定。采用Transwell小室法和Western blot检测TNC对细胞迁移、侵袭能力以及上皮间质转化(EMT)相关分子表达的影响。结果成功构建TNC质粒,稳定过表达TNC的PANC1细胞迁移、侵袭能力显著增强(P<0.05),上皮标志物E-cadherin表达减少而间质标志物N-cadherin表达增加(P<0.05)。结论TNC真核表达质粒以及TNC稳定表达胰腺癌细胞系的建立,可用于TNC对细胞EMT、迁移和侵袭等方面的机制研究,也是研究TNC生物学功能和胰腺癌发生发展机制的基础。Objective To construct human TNC expression plasmid for the investigation of the role of TNC in pancreatic cancer metastasis by using pancreatic cancer cell strain with stable over-expression of TNC.Methods TNC expression plasmid was constructed by unique primer design method,and then pancreatic cancer cell strain PANC1 with stable TNC expression were obtained by transfection and G418 screening.RT-qPCR and Western blot examined the expression of TNC.Transwell and Western blot were used to detecte the effects of TNC on cell migration,invasion and EMT-related molecules.Results TNC plasmid was successfully constructed.TNC promoted cell migration and invasion(P<0.05),decreased epithelial marker E-cadherin expression and upregulated mesenchymal marker N-cadherin expression(P<0.05).Conclusions The TNC expression plasmid and TNC stable expression pancreatic cancer cell strain may be used to explore the biological function of TNC and the pathogenesis of pancreatic cancer.

关 键 词：肌腱蛋白C 胰腺癌 迁移 侵袭 上皮间质转化 

分 类 号：R735.9[医药卫生—肿瘤]