Zhou Yun, Zhang Yan, Wang Jia
Department of Clinical Laboratory Medicine, Shanghai Tenth People's Hospital of Tongji University, Shanghai 200072, People's Republic of China.
Cancer Manag Res. 2020 Oct 29;12:10705-10713. doi: 10.2147/CMAR.S274578. eCollection 2020.
Pancreatic cancer (PC) is a malignant tumor with poor prognosis. This study aimed to determine the role of trefoil factor 2 (TFF2) in the proliferation and apoptosis of LPS-induced normal pancreatic duct cells and pancreatic cancer cells through β-catenin pathway.
TFF2 expression in normal pancreatic duct cells, pancreatic cancer cells and LPS-induced normal pancreatic duct cells was detected by RT-qPCR analysis and Western blot analysis. The transfection effects in pancreatic cancer cells and LPS-induced normal pancreatic duct cells were analyzed by RT-qPCR analysis. After indicated transfection, proliferation, apoptosis and inflammation of these cells were respectively detected by CCK-8 assay, TUNEL assay and certain ELISA kits. Expression of β-catenin pathway-related proteins was analyzed by Western blot analysis. Co-immunoprecipitation assay determined the combination of TFF2 and β-catenin.
TFF2 expression was increased in pancreatic cancer cells and LPS-induced HPDE cells compared with HPDE cells. According to TFF2 expression in these cells, PanC-1 cells and 5 μg/mL LPS were selected. In addition, TFF2 interference decreased the proliferation and promoted the apoptosis of PanC-1 cells and LPS-induced HPDE cells. However, TFF2 interference did not obviously change the levels of TNF-α, IL-1β and IL-6 in PanC-1 cells and LPS-induced HPDE cells. Furthermore, TFF2 interference suppressed the expression of β-catenin, c-Myc, Cyclin D1 and BIRC5 in PanC-1 cells and LPS-induced HPDE cells. TFF2 was demonstrated to combine with β-catenin.
TFF2 interference inhibits proliferation and promotes apoptosis of PanC-1 cells and LPS-induced HPDE cells by suppressing β-catenin pathway.
胰腺癌（PC）是一种预后较差的恶性肿瘤。本研究旨在通过β-连环蛋白途径确定三叶因子2（TFF2）在脂多糖（LPS）诱导的正常胰腺导管细胞和胰腺癌细胞增殖与凋亡中的作用。
通过逆转录定量聚合酶链反应（RT-qPCR）分析和蛋白质免疫印迹分析检测TFF2在正常胰腺导管细胞、胰腺癌细胞及LPS诱导的正常胰腺导管细胞中的表达。通过RT-qPCR分析评估胰腺癌细胞和LPS诱导的正常胰腺导管细胞中的转染效果。在进行指定转染后，分别采用细胞计数试剂盒（CCK-8）检测、末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）检测及特定酶联免疫吸附测定（ELISA）试剂盒检测这些细胞的增殖、凋亡和炎症情况。通过蛋白质免疫印迹分析β-连环蛋白途径相关蛋白的表达。免疫共沉淀实验确定TFF2与β-连环蛋白的结合情况。
与人胰腺导管上皮细胞（HPDE）相比，胰腺癌细胞及LPS诱导的HPDE细胞中TFF2表达升高。根据这些细胞中TFF2的表达情况，选择了胰腺癌细胞系PanC-1细胞及5μg/mL LPS。此外，TFF2干扰降低了PanC-1细胞及LPS诱导的HPDE细胞的增殖并促进其凋亡。然而，TFF2干扰并未明显改变PanC-1细胞及LPS诱导的HPDE细胞中肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）的水平。此外，TFF2干扰抑制了PanC-1细胞及LPS诱导的HPDE细胞中β-连环蛋白、c-Myc、细胞周期蛋白D1（Cyclin D1）和凋亡抑制蛋白5（BIRC5）的表达。证实TFF2与β-连环蛋白结合。
TFF2干扰通过抑制β-连环蛋白途径抑制PanC-1细胞及LPS诱导的HPDE细胞的增殖并促进其凋亡。
