EphA2蛋白调控鼻咽癌紫杉醇敏感性的实验研究

Mediation of paclitaxel sensitivity with EphA2 proteinin nasopharyngeal carcinoma in vitro

摘要:

目的 探讨促红细胞生成素产生肝细胞受体A2（erythropoietinproducing hepatocellular receptor, EphA2）蛋白对鼻咽癌细胞紫杉醇化疗敏感性的影响。方法 采用脂质体2000将EphA2蛋白过表达载体pEGFPN1EphA2和空白载体pEGFPN1分别转染鼻咽癌5-8F细胞，Western blot验证转染效果，CCK8法检测各组细胞生存率，并计算各组紫杉醇药物IC50值；流式细胞术检测紫杉醇作用后鼻咽癌5-8F细胞周期及凋亡率的改变。结果 鼻咽癌58F细胞EphA2蛋白上调后，紫杉醇药物在亲本细胞组和空白载体对照组中的IC50值分别为（1.4 ± 0.05）nM/L和（1.3 ± 0.06）nM/L，而实验组中的表达为（3.8 ± 0.52）nM/L，两组比较差异有统计学意义（P<0.05）。细胞周期结果显示实验组（45.76±3.89）%较亲本细胞组（65.85±2.28）%和对照组（64.52±3.31）%的G0/G1期细胞的比例明显减少，两组比较差异具有统计学意义（P<0.05）；而S期细胞的实验组（31.56±1.59）%较亲本细胞组（25.76±1.89）%和对照组（24.55±3.64）%和G2/M期细胞的实验组（23.10±4.55）% 较亲本细胞组（8.39±0.81）%和对照组（10.94±3.27）%的比例则明显增多，两组比较差异均具有统计学意义（P<0.05）；而细胞凋亡率3组之间比较差异无统计学意义（P>0.05）。结论 该研究结果表明EphA2蛋白能通过影响鼻咽癌细胞周期改变而调控其对紫杉醇药物的敏感性。 

Abstract:

Objective To investigate the effect of EphA2 protein on the chemosensitivity of paclitaxel in nasopharyngeal carcinoma (NPC) in vitro. Methods NPC 58F cells were transfected with EphA2 overexpression vector pEGFPN1EphA2 and empty vector pEGFPN1 by LipofectAMINE 2000 and Western blot was used for detecting the transfection efficiency. The IC50 values of paclitaxel in each group were obtained by CCK8 assays. Changes of cell cycle and apoptosis of 58F cells were examined with fluorescenceactivated cell sorting analysis. Results Compared with parental 58F cells (1.40±0.05) nM/L and 58F cells transfected with empty vector (1.30±0.06) nM/L, the IC50 value of paclitaxel was significantly upregulated in 58F cells transfected with EphA2 overexpression vector (3.80±0.52) nM/L (P<0.05). Meantime, cells in G0/G1 phase were greatly decreased ［(45.76±3.89)% vs (65.85±2.28)%, (45.76±3.89)% vs (64.52±3.31)%］, but cells in S phase ［(31.56±1.59)% vs (25.76±1.89)%, (31.56±1.59)% vs (24.55±3.64)%)］ and G2/M phase ［(23.10±4.55)% vs (8.39±0.81)%, (23.10±4.55)% vs (10.94±3.27%)］ were obviously increased (P<0.05). However, no significant difference was found in cell apoptosis rate among the 58F cells of the three groups (P>0.05). Conclusion EphA2 can regulate the chemosensitivity of paclitaxel via influencing the cell cycle in NPC cells.