CAJ | 학술논문

目的:以往研究已证实,棱果榕树叶的乙醇提取物及其乙酸乙酯萃取物(ethyl acetate soluble fraction,EASF)对人类乳腺癌细胞T47D有很强的细胞毒效应.本研究将进一步明确棱果榕树叶乙醇提取物的EASF联合阿霉素对人类乳腺癌T47D细胞系在细胞毒性、细胞周期阻滞以及诱导细胞凋亡方面的协同效应.方法:使用溴化噻唑蓝四氮唑比色法分析T47D细胞毒性效应,流式细胞仪进行细胞周期分析,ModFit LT3.0程序进行数据处理；采用溴化乙锭/吖啶橙双染色法检测细胞凋亡；免疫组织化学法识别T47D细胞系的聚ADP-核苷酸聚合酶(poly ADP-ribose polymerase,PARP)的表达.结果:EASF(0.875～7 μg/mL)与阿霉素(2～8 nmol/L)联合使用比单纯使用阿霉素能更有效地抑制T47D细胞生长.此外,二者联合使用能够增加细胞凋亡的发生率.研究发现,EASF能通过改变细胞从G2/M期至G1期而增强阿霉素的细胞毒效应.并且,二者的结合比单独使用能刺激T47D细胞中裂解性PARP的表达.结论:EASF可以通过诱导细胞凋亡和细胞周期停滞加强T47D细胞中的阿霉素活性.
목적:이왕연구이증실,릉과용수협적을순제취물급기을산을지췌취물(ethyl acetate soluble fraction,EASF)대인류유선암세포T47D유흔강적세포독효응.본연구장진일보명학릉과용수협을순제취물적EASF연합아매소대인류유선암T47D세포계재세포독성、세포주기조체이급유도세포조망방면적협동효응.방법:사용추화새서람사담서비색법분석T47D세포독성효응,류식세포의진행세포주기분석,ModFit LT3.0정서진행수거처리；채용추화을정/아정등쌍염색법검측세포조망；면역조직화학법식별T47D세포계적취ADP-핵감산취합매(poly ADP-ribose polymerase,PARP)적표체.결과:EASF(0.875～7 μg/mL)여아매소(2～8 nmol/L)연합사용비단순사용아매소능경유효지억제T47D세포생장.차외,이자연합사용능구증가세포조망적발생솔.연구발현,EASF능통과개변세포종G2/M기지G1기이증강아매소적세포독효응.병차,이자적결합비단독사용능자격T47D세포중렬해성PARP적표체.결론:EASF가이통과유도세포조망화세포주기정체가강T47D세포중적아매소활성.
OBJECTIVE:Previously,ethanolic extract of Ficus septica Burm.f.(Moraceae) leaves and its ethyl acetate soluble fraction (EASF) exhibited potent cytotoxic effects on T47D breast cancer cells.In the present study,we further investigated the effects of EASF of ethanolic extract of F.septica leaves in combination with doxorubicin on T47D breast cancer cell line in cytotoxicity,cell cycle arrest and apoptosis induction.METHODS:The cytotoxic effect analysis on T47D cells was carried out using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay.Analysis of cell cycle distribution was performed using a flowcytometer and the data were analyzed using ModFit LT 3.0 program.Apoptosis assay was carried out by double staining method using ethydium bromide-acridin orange.The expression of cleaved-poly ADP-ribose polymerase in the T47D cell line was identified using immunohistochemical techniques.RESULTS:The combination of doxorubicin (2 to 8 nmol/L) with EASF (0.875 to 7 μg/mL)more potently inhibited cell growth than the single treatment of doxorubicin in T47D cells.In addition,the combination of doxorubicin and EASF could increase the incidence of cells undergoing apoptosis.EASF was found to improve cytotoxic effect of doxorubicin by changing the inhibition of cell cycle G2/M to G1 phase.The combination also exhibited a more intensive stimulatory effect on cleaved-PARP expression in T47D cells than the single treatment.CONCLUSION:It is concluded that EASF may enhance doxorubicin activities in T47D cells by inducing apoptosis and cell cycle arrest.