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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,ModFitLT3.0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(0.875～7μg/mL)与阿霉素(2～8nmol/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。
OBJECTIVE: Previous studies have confirmed that the ethanol extract of E. citrifolia leaves and its ethyl acetate soluble fraction (EASF) have a strong cytotoxic effect on human breast cancer cell T47D. This study will further clarify the synergistic effect of EASF combined with doxorubicin on the human breast cancer T47D cell line in terms of cytotoxicity, cell cycle arrest, and induction of apoptosis. Methods: T47D cytotoxicity was analyzed by thiazolyl blue tetrazolium bromide assay, cell cycle analysis was performed by flow cytometry, data processing was performed by ModFitLT3.0 program, and cells were detected by ethidium bromide/acridine orange double staining method. Apoptosis; Immunohistochemical identification of poly ADP-ribose polymerase (PARP) expression in T47D cell line. RESULTS: The combination of EASF (0.875-7 μg/mL) and adriamycin (2-8 nmol/L) was more effective than doxorubicin alone in inhibiting T47D cell growth. In addition, the combination of the two can increase the incidence of apoptosis. Studies have found that EASF can enhance the cytotoxic effect of doxorubicin by changing cells from the G2/M phase to the G1 phase. Also, the combination of the two can stimulate the expression of lytic PARP in T47D cells than when used alone. CONCLUSION: EASF can enhance doxorubicin activity in T47D cells by inducing apoptosis and cell cycle arrest.