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目的探讨促红细胞生成素产生肝细胞受体A2(erythropoietin-producing hepatocellular re-ceptor,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(2 5.7 6±1.8 9)%和对照组(2 4.5 5±3.6 4)%和G2/M期细胞的实验组(2 3.1 0±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。
Objective To investigate the effect of erythropoietin-producing hepatocellular re-ceptor (EphA2) protein on chemosensitivity to paclitaxel in nasopharyngeal carcinoma cells. METHODS: EphA2 overexpression vector pEGFP-N1-EphA2 and blank vector pEGFP-N1 were transfected into nasopharyngeal carcinoma 5-8F cells by lipofectamine 2000. The transfection efficiency was verified by Western blot. The cell viability was detected by CCK-8 assay The IC50 values of paclitaxel in each group were calculated. The cell cycle and apoptosis rate of 5-8F in nasopharyngeal carcinoma were detected by flow cytometry. Results The IC50 values of paclitaxel drug in the parent cell group and the blank vector control group were (1.4 ± 0.05) nM / L and (1.3 ± 0.06) nM / L respectively after the up-regulation of EphA2 protein in nasopharyngeal carcinoma 5-8F cells. The expression in the group was (3.8 ± 0.52) nM / L, the difference between the two groups was statistically significant (P <0.05). The results of cell cycle showed that the percentage of G0 / G1 phase cells in the experimental group (45.76 ± 3.89)% was significantly lower than that in the parental cell group (65.85 ± 2.28)% and the control group (64.52 ± 3.31)%, the difference was statistically significant (31.56 ± 1.59)% compared with the parental cell group (2 5.7 6 ± 1.8 9)% and the control group (4.55 ± 3.6 4)% and the G2 / M phase cell (P <0.05) (2 3.1 ± 4.55)% in the experimental group was significantly higher than that in the parental cell group (8.39 ± 0.81)% and the control group (10.94 ± 3.27)%, the difference between the two groups was statistically significant (P0.05) ; While the apoptosis rate of the three groups showed no significant difference (P> 0.05). Conclusion The results of this study indicate that EphA2 protein regulates its sensitivity to paclitaxel by affecting the cell cycle of NPC cells.