CAJ | 학술논문

目的初步探讨葛根总黄酮(PRF)对骨髓瘤细胞株U266及RPMI 8226增殖影响及其机制.方法采用0、10、30、50、100 μg/ml PRF分别处理U266、RPMI 8226细胞48h及72 h,MTT法检测细胞增殖抑制率,流式细胞术检测细胞周期变化,瑞特染色观察细胞形态学改变,FITC-Annexin V/PI双染法检测细胞早期凋亡率改变,DNA片段化分析观察PRF处理U266细胞DNA断裂片段.结果 PRF可以抑制2种骨髓瘤细胞增殖,对U266细胞抑制作用大于对RPMI 8226细胞的作用,呈浓度依赖性；随PRF浓度增加2种细胞凋亡比例增加.瑞特染色未观察到2种细胞凋亡形态学特征.0、10、30、50、100 μg/ml PRF处理U266细胞48 h早期凋亡率分别为(3.20±0.36)％、(5.20±0.92)％、(7.30±1.22)％、(8.10±0.53)％、(10.80±0.90)％,呈剂量依赖性增高,组间差异有统计学意义(P＜0.05).DNA片段化分析U266细胞未出现凋亡细胞特有的DNA梯带.结论一定浓度PRF对骨髓瘤U266及RPMI 8226细胞具有较明显的增殖抑制作用；PRF对U266细胞增殖抑制作用机制虽可能与凋亡相关,但并非U266细胞增殖受抑的主要途径,其确切机制有待进一步探讨.
목적 초보탐토갈근총황동(PRF)대골수류세포주U266급RPMI 8226증식영향급기궤제.방법 채용0、10、30、50、100 μg/ml PRF분별처리U266、RPMI 8226세포48h급72 h,MTT법검측세포증식억제솔,류식세포술검측세포주기변화,서특염색관찰세포형태학개변,FITC-Annexin V/PI쌍염법검측세포조기조망솔개변,DNA편단화분석관찰PRF처리U266세포DNA단렬편단.결과 PRF가이억제2충골수류세포증식,대U266세포억제작용대우대RPMI 8226세포적작용,정농도의뢰성；수PRF농도증가2충세포조망비례증가.서특염색미관찰도2충세포조망형태학특정.0、10、30、50、100 μg/ml PRF처리U266세포48 h조기조망솔분별위(3.20±0.36)％、(5.20±0.92)％、(7.30±1.22)％、(8.10±0.53)％、(10.80±0.90)％,정제량의뢰성증고,조간차이유통계학의의(P＜0.05).DNA편단화분석U266세포미출현조망세포특유적DNA제대.결론 일정농도PRF대골수류U266급RPMI 8226세포구유교명현적증식억제작용；PRF대U266세포증식억제작용궤제수가능여조망상관,단병비U266세포증식수억적주요도경,기학절궤제유대진일보탐토.
Objective To investigate the effects on proliferation of multiple myeloma cell lines U266 and RPMI 8226 induced by puerariae radix flavones (PRF) in vitro and its possible mechanism.Methods Exposed to 0,10,30,50,100 μg/ml PRF for 48 h and 72 h,the U266 and RPMI 8226 cells proliferation inhibitory rates were detected by MTT assay,cell cycles by flow cytometry (FCM),morphologic changes of U266 cells by Wright' s staining,and early-stage apoptotic rates of U266 cells by FITC-Annexin V/PI staining with FCM.Analysis of DNA fragment was made to test characteristic apoptosis DNA ladder in U266 cells.Results 0,10,30,50,100 μg/ml PRF could inhibit the proliferation of U266 and RPMI 8226 cells in a dose-dependent manner (U266 ＞ RPMI 8226).Cell cycle analyses in U266 and RPMI 8226 cells showed that sub-diploid peaks,but cell cycles changed minor.Wright's staining of U266 cells showed hardly any apoptostic character istic.Annexin V/PI double staining indicated that early-stage apoptotic rates of U266 cells exposed to 0,10,30,50,100 μg/ml PRF for 48 h were mildly increased in a dose-dependent manner.They were (3.20±0.36) ％,(5.20±0.92) ％,(7.30±1.22) ％,(8.10±0.53) ％ and (10.80±0.90) ％,respectively.The group differences had statistical significance (P ＜ 0.05).Analysis of DNA fragment barely exhibited the characteristic DNA ladder in U266 cells.Conclusion A certain concentrations of PRF could inhibit the proliferation of U266 and RPMI 8226 cells significantly.It is suggested that apoptosis related to the proliferative inhibition mechanism induced by PRF in U266 cell line,but not main.Other pathways such as necrosis and autophagy whether or not involved need further investigation.