论文部分内容阅读
目的探讨1,2-DCE诱导人骨髓神经母细胞瘤细胞株SH-SY5Y细胞凋亡及对凋亡相关蛋白表达的影响。方法以终浓度分别为0、10、20、30、40、50、60、70和80 mmol/L的1,2-DCE染毒SH-SY5Y细胞,培养24 h后,采用磷脂结合蛋白Ⅴ-异硫氰酸荧光素/碘化丙啶流式细胞技术检测细胞的凋亡情况,采用免疫印迹法检测细胞中P53、B细胞淋巴瘤/白血病-2(BCL-2)和BCL-2相关X蛋白(BAX)的蛋白相对表达水平。结果在浓度为0~80mmol/L时,随着1,2-DCE染毒剂量的增加,SH-SY5Y细胞总凋亡率升高,呈剂量-效应关系(P<0.01);与对照组比较,在1,2-DCE染毒剂量为30~80 mmol/L时,SH-SY5Y细胞早期凋亡率和总凋亡率均出现有统计学意义的升高(P<0.05)。与其余各组比较,P53蛋白相对表达水平在1,2-DCE染毒剂量为20 mmol/L时最低(P<0.05),BCL-2蛋白相对表达水平和BCL-2/BAX比值均在1,2-DCE染毒剂量为70 mmol/L时最低(P<0.05);但未见P53、BCL-2、BAX的蛋白相对表达水平及BCL-2/BAX比值与1,2-DCE染毒剂量之间呈现有规律的剂量依赖性改变。多元线性回归分析结果显示,1,2-DCE染毒后,SH-SY5Y细胞总凋亡率与P53、BCL-2的蛋白相对表达水平以及BCL-2/BAX比值均存在关联(P<0.05)。结论 1,2-DCE可导致SH-SY5Y细胞凋亡,其机制可能与P53、BCL-2蛋白表达及BCL-2/BAX相对量的改变有关。
Objective To investigate the effects of 1,2-DCE on the apoptosis of SH-SY5Y cells and the expression of apoptosis-related proteins in human myeloblastoma cell line SH-SY5Y. Methods SH-SY5Y cells were treated with 1,2-DCE at final concentrations of 0, 10, 20, 30, 40, 50, 60, 70 and 80 mmol / L, Fluorescein isothiocyanate / propidium iodide flow cytometry was used to detect cell apoptosis. Western blotting was used to detect the expression of P53, B-cell lymphoma / leukemia-2 (BCL-2) and BCL-2 Protein (BAX) protein relative expression level. Results The total apoptotic rate of SH-SY5Y cells increased with dose of 1,2-DCE at a concentration of 0-80 mmol / L, showing dose-effect relationship (P <0.01). Compared with the control group The apoptotic rate and the total apoptosis rate of SH-SY5Y cells were significantly increased at doses of 30-80 mmol / L 1,2-DCE (P <0.05). Compared with the other groups, the relative expression level of P53 protein was the lowest (P <0.05) at the dose of 1,2-DCE of 20 mmol / L, and the relative expression level of BCL-2 protein and BCL-2 / BAX ratio were all 1 (P <0.05). However, the relative expression levels of P53, BCL-2 and BAX and the ratio of BCL-2 / BAX to 1,2-DCE were not significantly different The dose showed a regular dose-dependent changes. The results of multiple linear regression analysis showed that there was a significant correlation between the total apoptotic rate of SH-SY5Y cells and the relative expression levels of P53 and BCL-2 and the ratio of BCL-2 / BAX after 1,2-DCE exposure (P <0.05) . Conclusions 1,2-DCE can induce the apoptosis of SH-SY5Y cells, which may be related to the changes of P53 and BCL-2 protein expression and the relative amount of BCL-2 / BAX.