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目的:探讨南蛇藤茎乙酸乙酯提取物(Celastrus orbiculatus extract,COE)抗人胶质母细胞瘤U251细胞株的增殖和凋亡作用,研究COE抗肿瘤的分子机制,为寻找新的治疗胶质母细胞瘤药物提供依据。方法:以人胶质母细胞瘤细胞株U251为研究对象,设空白组和COE组,应用噻唑蓝[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]比色法观察COE对细胞活力的抑制作用;5-溴脱氧尿嘧啶核苷(5-bromo-2-deoxy uridine,Brd U)标记检测COE对U251细胞增殖的影响;Annexin V/碘化丙啶(PI)双标法检测COE对U251细胞凋亡的作用;透射电镜下观察经COE干预后U251细胞超微结构变化;蛋白免疫印迹法(Western blot)检测COE对凋亡标志基因B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2),Bcl-2相关X蛋白(Bcl-2-associated X,Bax)和半胱氨酸蛋白酶-3(Caspase-3)蛋白表达的影响。结果:与空白组比较,COE能够明显抑制U251细胞的增殖(P<0.05);诱导U251细胞的早期凋亡,电镜下可见到凋亡小体,影响并改变U251细胞超微结构;COE能促进Caspase-3,Bax蛋白表达,降低Bcl-2,Bcl-2/Bax蛋白表达(P<0.05)。结论:COE能有效抑制胶质母细胞瘤U251细胞株的增殖,促进肿瘤细胞凋亡,其作用机制与调节Bcl-2和Caspase-3表达有关。
OBJECTIVE: To investigate the proliferation and apoptosis of Celastrus orbiculatus extract (COE) -treated human glioblastoma U251 cell line, to study the molecular mechanism of COE antitumor and to find a new therapeutic glue Glioblastoma drug provides the basis. Methods: The human glioblastoma cell line U251 was used as the research object. The blank group and the COE group were established. The cells were treated with 3 - (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2-H (MTT) assay was used to observe the inhibitory effect of COE on cell viability. The effect of COE on the proliferation of U251 cells was detected by 5-bromo-2-deoxy uridine (5-bromo-2-deoxy uridine) The effect of COE on the apoptosis of U251 cells was detected by V / propidium iodide (PI) double staining. The ultrastructural changes of U251 cells were observed under transmission electron microscope. The expressions of COE on apoptosis were detected by Western blot. Bcl-2, Bcl-2, Caspase-3, Bcl-2, Effect of protein expression. RESULTS: Compared with the blank group, COE significantly inhibited the proliferation of U251 cells (P <0.05); induced the early apoptosis of U251 cells, which could be seen by electron microscopy, which affected and changed the ultrastructure of U251 cells; COE promoted Caspase-3, Bax protein expression, Bcl-2, Bcl-2 / Bax protein expression (P <0.05). CONCLUSION: COE can effectively inhibit the proliferation of glioblastoma U251 cell line and promote the apoptosis of tumor cells. Its mechanism is related to the regulation of Bcl-2 and Caspase-3 expression.