广西虎纹捕鸟蛛毒诱导U251胶质瘤细胞凋亡 及Caspase-3的激活

Research on Induction of Apoptosis of Neurogliocytoma U251 and Activation of Caspase-3 by Toxin of Selenocosmia huwena in Guangxi

摘要:

目的: 探讨虎纹捕鸟蛛毒抑制体外脑胶质瘤细胞系U251生长并诱导其凋亡,激活半胱天冬酶-3(Caspase -3)的作用。 方法: 将U251细胞分为3组:空白组、顺铂组、加药组(虎纹捕鸟蛛毒素浓度为37.5,50,75,100,150 mg ·L^-1),毒素作用24,48 h后用四甲基偶氮唑蓝(MTT )法检测细胞活性并与顺铂组、空白组做对比;流式细胞仪AnnexinV-FITC检测细胞凋亡率;Caspase-3分光光度法检测Caspase-3的相对活性。 结果: 虎纹捕鸟蛛毒素剂量为37.5,50,75,100,150 mg ·L^-1时,对U251细胞的增殖有显著性抑制作用(P<0.05或P<0.01),IC₅₀为53.48 mg ·L^-1。虎纹捕鸟蛛毒素可诱导U251胶质瘤细胞凋亡并呈浓度依赖关系。在剂量为150 mg ·L^-1诱导48 h,细胞早期凋亡率84.1%,晚期凋亡率4.48%。用不同浓度分别处理细胞,Caspase-3活性于48 h,150 mg ·L^-1时达高峰,对细胞中的执行分子Caspase-3的激活速度快,活化程度为9.23,并存在剂量依赖关系和浓度依赖性的升高。 结论: 虎纹捕鸟蛛毒素明显抑制U251细胞生长并诱导其发生凋亡, 凋亡过程中有Caspase-3的激活。

Abstract:

Objective: To explore the in vitro effect of the toxin of Selenocosmia huwena on inhibiting the growth and inducing the death of cells of neurogliocytoma, and activating caspase-3. Method: MTT was applied to obtain the growth curves of cell affected by different concentrations of toxin of S.huwena for 24 h and 48 h. AnnexinV-FITC was used to test the death rate. Absorption spectrometry was adopted to measure the relative activity of caspase-3. Result: The toxin of S.huwena evidently inhibited the growth of U251, and it could induce the death of cells neurogliocytoma with concentration-dependent manner. With cells affected by different concentrations of toxin for 24 h, we found that the activity of caspase-3 reached the maximum at 48 h with concentration-dependent manner. Conclusion: The toxin of S.huwena evidently inhibits the growth of U251 and induces the death of U251, and the process include the activation of caspase-3.