乳腺癌细胞凋亡中放线菌素D作用的研究

Study on breast carcinoma cells apoptosis induced by Act-D

目的探讨热休克蛋白(HSP)在Act—D诱导乳腺癌MDA-MB-435s细胞凋亡中的作用,为进一步探讨HSP抗凋亡机制及乳腺癌生物治疗提供实验依据。方法体外培养的乳腺癌MDA-MB-435s细胞于RPMI1640、10%胎牛血清、5%CO2孵育,取指数生长期细胞1×105/ml接种反应板24 h,分为非热休克组(NHS)和热休克组(HS),两组分别由不同浓度(0,5,10,20μg/ml)放线菌素D(Act—D)诱导,作用4 h。采用脱氧核苷酸末端转移酶介导的dUTP缺口末端标记法(TUNEL)和3H-胸腺嘧啶脱氧核苷(3H-TdR)掺入试验检测肿瘤细胞凋亡指数(AI)及DNA合成情况。结果 HS组比NHS组凋亡指数明显下降(P<0.01);cpm值明显升高(P<0.01),呈现剂量依赖效应。结论热休克反应对乳腺癌细胞经Act-D诱导凋亡具有抑制作用,此作用与Act-D呈剂量依赖效应。

Objective To investigate the effect of HSP of breast carcinoma cells MDA-MB-435s apoptosis induced by Actinomycin-D.Methods breast carcinoma cells which are cultured in vitro are grouped NHS and HS groups before treated by Act-D.TUNEL is used to exam apoptosis index and 3H-TdR test is used to examing DNA synthsis.Results The apoptosis index of HS group is more significant decreased than the one of NHS group（P0.01）.And the cpm of HS group is more significant increase than the one of NHS group（P0.01）,which are depended on the dosage of Act-D.Conclusion This paper results indicate that HSP has significant inhibition of apoptosis induced by Act-D in breast carcinoma cells in vitro.