月腺大戟素A抗乳腺癌活性

Anti-breast cancer activity of ebracteolatain A

目的从中药狼毒药材中分离、纯化化合物月腺大戟素A,探讨其抗乳腺癌活性。方法采用回流提取、溶剂萃取和吸附色谱法从狼毒药材中分离、纯化月腺大戟素A,通过质谱和核磁共振谱解析其化学结构。以乳腺癌细胞SUM149(三阴型)、MCF-7(luminal A型)、ZR-75-1(luminal B型)、SK-BR-3(HER2阳性型)为测试细胞株,用MTT法测定月腺大戟素A的抗乳腺癌细胞增殖活性,用流式细胞术检测月腺大戟素A对SUM149细胞周期的影响,用裸鼠移植SUM149细胞建立肿瘤模型并评价月腺大戟素A对肿瘤的抑制效果。结果月腺大戟素A结构为3,3’-二乙酰基-2,4’-二甲氧基-2’,4,6,6’-四羟基-5’-甲基二苯基甲烷;对SUM149、MCF-7、ZR-75-1和SK-BR-3细胞的半数抑制浓度分别为5.50、6.16、7.08、8.64μmol/L。月腺大戟素A作用SUM149细胞12、24和48 h后,随着月腺大戟素A浓度(2.5、5、10μmol/L)的升高,G0/G1期细胞比例下降(P<0.05,P<0.01),S期细胞比例上升(P<0.05,P<0.01)。给予35 mg/kg腹腔注射月腺大戟素A后,裸鼠肿瘤体积和质量的抑制率分别为37.94%和41.38%。结论月腺大戟素A对4种乳腺癌细胞增殖均有抑制作用,并可抑制裸鼠SUM149细胞移植肿瘤生长,其机制可能与抑制SUM149细胞周期从S期到G2/M期的转换有关。

Objective To isolate and purify ebracteolatain A from the roots of Euphorbia ebracteolata Hayata, and to explore its anti-breast cancer activity. Methods Ebracteolatain A was isolated and purified from the roots of Euphorbia ebracteolata Hayata using reflux extraction, solvent extraction and absorption chromatography techniques. The chemical structure was identified by mass spectrometry and nuclear magnetic resonance spectrometry. The antibreast cancer activity of ebracteolatain A was determined by MTT assay in the breast cancer cell lines including SUM149（triple-negative）, MCF-7（luminal A）, ZR-75-1（luminal B） and SK-BR-3（HER2-positive）. The effect of ebracteolatain A on cell cycle was determined by flow cytometry. A tumor model was established in nude mice by transplanting SUM149 cells, and the inhibitory effect of ebracteolatain A on breast cancer was evaluated. Results The ebracteolatain A was 3, 3’-diacetyl-2, 4’-dimethoxy-2’, 4, 6, 6’-tetrahydroxy-5’-methyl diphenylmethane. The half maximal inhibitory concentrations of ebracteolatain A on SUM149, MCF-7, ZR-75-1 and SK-BR-3 cells were 5.50, 6.16, 7.08 and 8.64 μmol/L, respectively. With the increase of drug concentration（2.5, 5, 10 μmol/L）, the percentage of the cells at G0/G1 phase was decreased（P〈0.05, P〈0.01） and the percentage of the cells at S phase was increased（P〈0.05, P〈0.01） after treatment with ebracteolatain A for 12, 24 and 48 h. After intraperitoneal injection of ebracteolatain A 35 mg/kg, the inhibition rates of the tumor volume and mass in nude mice were 37.94% and 41.38%, respectively. Conclusion Ebracteolatain A can inhibit the proliferation of the four types of breast cancer cells and the growth of transplanted-SUM149 cell tumor on nude mice, which may be related to suppressing the transition of cell cycle from S phase to G2/M phase.