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目的:检测不同类型肿瘤化疗药物对视网膜母细胞瘤(retinoblastoma RB)细胞系 HXO-RB_(44) 的诱导凋亡作用,建立该细胞系的细胞凋亡模型,作为今后研究化疗药物诱导RB细胞凋亡机理的工作基础。方法:将长春新碱、阿糖胞苷、氨甲喋呤、环磷酰胺、噻替派、米托蒽醌、阿克拉霉素、吡喃阿霉素、顺铂、卡铂、丝裂霉素、足叶乙甙等十二种化疗药物分别以10~(-9) 、10~(-8)、10~(-7)、10~(-6)、10~(-5)、10~(-4)mol·L~(-1)的浓度加入RB细胞中培养24小时,利用DNA凝胶电泳技术,检测RB细胞在不同化疗药物、不同浓度作用下的凋亡情况;将其中能产生明显凋亡梯带的化疗药物按最适浓度加入RB细胞中分别培养4、8、16、24、48、72小时,检测在不同时间的作用下,HXO-RB_(44)细胞系的细胞凋亡现象,并运用透射电子显微镜技术从形态学上观察凋亡细胞的存在。结果:10~(-6)～10~(-5)mol·L~(-1)长春新碱及10~(-5)mol·L~(-1)阿克拉霉素作用于HXO-RB_(44)细胞系24小时后,DNA凝胶电泳出现典型的限制性降解片段梯带,其余十种化疗药物作用后均未见该特征性表现。取10~(-5)mol·L~(-1)长春新碱作用于RB细胞,8小时开始出现DNA电泳梯带,24小时最明显,48小时后减弱。透射电镜可观察到大量具有明显形态学特征的凋亡RB细胞。结论:长春新碱和?
OBJECTIVE: To detect the apoptosis induced by different types of tumor chemotherapeutic drugs on retinoblastoma RB cell line HXO-RB_ (44), and to establish a cell apoptosis model of this cell line as a basis for future study of chemotherapeutic drug-induced RB cell apoptosis Work mechanism of death mechanism. Methods: Vincristine, cytarabine, methotrexate, cyclophosphamide, thiotepa, mitoxantrone, aclacinomycin, pirarubicin, cisplatin, carboplatin, mitomycin, Etoposide and other 12 chemotherapeutic drugs were administered orally at 10 ~ (-9), 10 ~ (-8), 10 ~ (-7), 10 ~ (-6), 10 ~ (-5), 10 ~ 4) at a concentration of mol·L -1 for 48 hours. The apoptosis of RB cells under different chemotherapeutic agents and different concentrations was detected by DNA gel electrophoresis. The apoptosis-inducing chemotherapeutic drugs were added to RB cells at the optimal concentration for 4, 8, 16, 24, 48 and 72 hours respectively to detect the apoptosis of HXO-RB_ (44) cell lines under different time. The morphology of apoptotic cells was observed by transmission electron microscopy. Results: Vincristin and 10-5 mol·L -1 aclacinomycin reacted with 10 -6 to 10 -5 mol·L -1 of HXO-RB_ (44) After 24 hours, typical restriction fragment degradation bands appeared on DNA gel electrophoresis, and none of the other 10 chemotherapeutic drugs showed this characteristic expression. Vincristine (10 -5 mol·L -1) was applied to RB cells. DNA ladder began to appear at 8 hours, which was the most obvious at 24 hours and weakened after 48 hours. A large number of apoptotic RB cells with obvious morphological features were observed by TEM. Conclusion: Vincristine and?