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目的 :探讨潜伏膜蛋白1(latent membrane protein 1,LMP1)瞬时过表达对鼻咽癌CNE2细胞迁移的影响及其可能的作用机制。方法 :成功构建重组真核表达载体pEGFP-C1-LMP1,并将其瞬时转染至CNE2细胞,激光共聚焦显微镜下观察LMP1在CNE2细胞中的定位,Transwell法检测LMP1对CNE2细胞迁移和侵袭能力的影响,实时荧光定量PCR和蛋白质印迹法检测LMP1和Ras相关C3肉毒素底物1(Ras-related C3 botulinum toxin substrate 1,Rac1)mRNA及蛋白的表达。结果 :pEGFP-C1-LMP1成功转染至CNE2细胞后,可见LMP1主要表达于细胞膜和部分细胞质中,CNE2细胞的侵袭和迁移能力明显低于转染空载体的对照组(P<0.05),CNE2细胞中LMP1 mRNA及蛋白的表达水平明显高于转染空载体的对照组(P<0.01),而Rac1 mRNA及蛋白的表达水平明显低于转染空载体的对照组(P<0.05)。结论 :LMP1瞬时过表达可抑制鼻咽癌CNE2细胞的迁移,这一作用可能与Rac1表达下调有关。
Objective: To investigate the effect of transient membrane protein 1 (LMP1) overexpression on the migration of nasopharyngeal carcinoma CNE2 cells and its possible mechanism. Methods: The recombinant eukaryotic expression vector pEGFP-C1-LMP1 was successfully constructed and transfected into CNE2 cells transiently. The localization of LMP1 in CNE2 cells was observed by laser scanning confocal microscopy. The migration and invasion ability of LMP1 on CNE2 cells were detected by Transwell assay The mRNA and protein expressions of LMP1 and Ras-related C3 botulinum toxin substrate 1 (Rac1) were detected by real-time fluorescence quantitative PCR and Western blotting. RESULTS: After transfection of pEGFP-C1-LMP1 into CNE2 cells, LMP1 was mainly expressed in the cell membrane and some cytoplasm. The invasion and migration ability of CNE2 cells was significantly lower than that of the control group transfected with empty vector (P <0.05). CNE2 The expression level of LMP1 mRNA and protein in cells was significantly higher than that in control cells transfected with empty vector (P <0.01), while the expression of Rac1 mRNA and protein was significantly lower than that in control group transfected with empty vector (P <0.05). CONCLUSION: Transient overexpression of LMP1 can inhibit the migration of nasopharyngeal carcinoma CNE2 cells, which may be related to the down-regulation of Rac1 expression.