Objective: To investigate the effect of ERK1/2 inhibitor AZD8330 on human Burkitt's lymphoma cell line Raji cells and its mechanism.
Methods: Raji cells were treated with different concentrations of AZD8330. CCK-8 was used to detect the cell viability. The apoptosis rate of Raji cells was detected by flow cytometry using Annexin V/PI-staining. Real-time PCR was used to assess the expression of Bcl-2, Bcl-xl, caspase-3 and VEGF genes. The protein expression level of Bcl-2, Bcl-xl, caspase-3 and p-ERK1/2 was tested with Western blot.
Results: The cell survival rate decreased to(62.09±0.86)%,(50.06±1.33)% and (39.13±2.34)% respectively after cells were treated with AZD8330 at 1.00 μmol/L in vitro for 24 h, 48 h and 72 h, and statistically significant differences were observed in groups with different time of treatment(P<0.05). Apoptosis of cells treated with AZD8330 at 0.10, 1.00, 10.00 μmol/L in vitro for 24 h, 48 h and 72 h was analyzed, and the statistically significant differences were observed in groups of different time and concentration treatment (P<0.05). AZD8330 induced Raji cell apoptosis and upregulated expression of Bcl-2, Bcl-xl, VEFG and decreased the expression of caspase-3 in a dose and time dependent manner, and statistically significant differences were observed in groups of different time and concentration treatment (P<0.05). At the same time, the Bcl-2, Bcl-xl and p-ERK1/2 proteins expression is suppressed obviously, but the expression of caspase-3 protein increased.
Conclusion: AZD8330 induces cell apoptosis by down-regulating the activation of ERK1/2 signal transduction pathway in Burkitt's lymphoma cell line Raji cells in a dose and time dependent manner.
目的: 探讨细胞外信号调节激酶1/2（ERK1/2）抑制剂AZD8330对Burkitt淋巴瘤细胞株Raji细胞的作用及其机制。
方法: Raji细胞用不同浓度的AZD8330进行处理；采用CCK-8检测细胞存活率；流式细胞术检测细胞凋亡情况；实时定量PCR法检测Bcl-2、Bcl-xl、caspase-3和血管内皮生长因子(VEGF）mRNA表达；Western blot法检测Bcl-2、Bcl-xl、caspase-3、磷酸化(p)-ERK1/2蛋白表达。
结果: 1.00 µmol/L的AZD8330处理24、48和72 h后细胞存活率分别为(62.09±0.86）%、(50.06±1.33）%和(39.13±2.34）%，差异有统计学意义（P值均<0.05）；0.10、1.00、10.00 µmol/L的AZD8330分别处理Raji细胞24、48和72 h，Raji细胞发生凋亡，凋亡率呈时间和剂量依赖性，差异有统计学意义（P值均<0.05）；随着浓度增加和时间延长，Bcl-2、Bcl-xl、VEGF mRNA表达降低，caspase-3 mRNA表达升高，差异有统计学意义（P值均<0.05）；同时，Bcl-2、Bcl-xl、p-ERK1/2蛋白表达明显受抑制，而caspase-3蛋白表达增强。
结论: AZD8330可能通过抑制ERK1/2通路相关基因和蛋白的表达而诱导Burkitt淋巴瘤Raji细胞凋亡，抑制其增殖。