前梯度蛋白2干扰质粒的构建及其对鼻咽癌细胞生物学功能的影响

摘要

AIM: To construct the shRNA targeting anterior gradient protein 2 (AGR2) gene for exploring the effect of AGR2 on the biological behavior of nasopharyngeal carcinoma (NPC) cells.METHODS: The expression of AGR2 at mRNA and protein levels in NPC cell lines 6-10B and 5-8F was detected by real-time PCR and Western blot.The pSR-GFP/Neo-AGR2-shRNA expression vector targeting AGR2 was constructed.Based on the interference targeting AGR2, the cell migration and motility were determined by Transwell migration and motility assays.RESULTS: The expression of AGR2 was increased in NPC cell line 5-8F compared with NPC cell line 6-10B (P <0.05).When the AGR2 expression in 5-8F cells was interfered, the cell migration, invasion and tumorigenicity were weakened.CONCLUSION: The expres-sion of AGR2 is up-regulated in NPC cell line 5-8F.pSR-GFP/Neo-CLU-shRNA successfully inhibits the expression of AGR2 in NPC cell line 5-8F.AGR2 inhibits the migration, invasion and tumorigenicity of 5-8F cells in vivo.%目的：构建前梯度蛋白2（AGR2）基因干扰重组质粒及建立 AGR2基因沉默的鼻咽癌细胞及动物模型，检测 AGR2对鼻咽癌细胞的生物学功能的影响。方法：首先以 real－time PCR 和 Western blot 验证 AGR2在鼻咽癌细胞系5－8F 和6－10B 中的表达，以高表达 AGR2的5－8F 细胞作为干扰转染的目标细胞；设计及构建 shRNA 表达载体 pSR－GFP／Neo－AGR2－shRNA，将其转染至5－8F 鼻咽癌细胞系中，通过 Transwell 小室迁移及侵袭实验来检测其迁移和侵袭功能的变化。结果：与6－10B 细胞比较，AGR2在5－8F 中的表达显著上调（P ＜0．05）；干扰鼻咽癌细胞5－8F 中的 AGR2可使细胞迁移和侵袭功能明显减弱（P ＜0．05），使其体外成瘤能力显著降低（P ＜0．05）。结论：AGR2在鼻咽癌细胞系5－8F 中明显高表达；pSR－GFP／Neo－AGR2－shRNA 可成功干扰5－8F 中 AGR2的表达；AGR2可抑制5－8F 细胞的迁移、侵袭及体内成瘤能力。