过表达的人β防御素-3可抑制结肠癌细胞增殖、迁移和细胞周期

Overexpression of hBD3 inhibits cell proliferation,cell cycle,and migration in colon cancer cells

本文研究探讨了人β防御素-3(hBD3)对结肠癌HCT116细胞的增殖、迁移和细胞周期的影响。将真核表达载体(pcDNA3.1-hBD3)转染于人结肠癌HCT116细胞。采用qPCR和Western blot方法检测转染效率。使用Western blot方法检测细胞中β-catenin,E-cadherin,N-cadherin蛋白的分子表达水平;采用单通道Hoechst染色检测细胞核数量和增殖能力;流式细胞仪检测细胞周期;划痕法和Transwell法检测结肠癌HCT116细胞的迁移能力。结果显示,转染过真核表达载体(pcDNA3.1-hBD3)的结肠癌HCT116细胞,hBD3的mRNA表达和蛋白表达水平显著高于结肠癌HCT116和转染pCMV-Blank的HCT116细胞;其增殖能力和迁移侵袭能力均受到不同程度的抑制;细胞周期的G2/M期受到阻滞;HCT116-hBD3细胞中β-catenin和N-cadherin两种蛋白的表达水平均低于HCT116和HCT116-Blank细胞,而E-cadherin蛋白表达的水平均高于HCT116和HCT116-Blank细胞。本文发现,hBD3或许是通过调控Wnt/β-catenin信号通路来调控结肠癌细胞HCT116的增殖、迁移和细胞周期。

In the present study,we investigated the effects of humanβ-defensin-3(hBD3)on the proliferation,cell cycle,and migration of HCT116 cells.The eukaryotic expression vector pcDNA3.1-hBD3 was transfected into human colon cancer HCT116 cells.The transfection efficiency was detected by qPCR and Western blotting analysis.The expressions ofβ-catenin,E-cadherin,and N-cadherin at the protein level were assessed by Western blotting analysis.Single-channel Hoechst33342 was used to detect the number of nuclei and calculate the proportion of proliferating cells.The cell cycle was detected by flow cytometry.The migration ability of HCT116 cells was detected by wound-healing and transwell assays.The results showed that the ability of migration and proliferation was inhibited,and the cell cycle G2/M phase was blocked.The expressions ofβ-catenin and N-cadherin at the protein level in HCT116-hBD3 cells were lower than those in HCT116 and HCT116-Blank cells,while the expression of E-cadherin at the protein level in HCT116-hBD3 cells was higher than that in HCT116 and HCT116-Blank cells.These findings suggested that hBD3 could regulate the migration and proliferation of HCT116 cells by regulating the Wnt/β-catenin signaling pathway.