Luz Lívia Eidam Camargo, Kanunfre Carla Cristine, Paludo Katia Sabrina, da Silva Justo Aline, Petry Victor Kubaski, Lemes Bruna Mikulis, Barison Andersson, Nepel Angelita, Wang Mei, Avula Bharathi, Khan Ikhlas Ahmad, Beltrame Flávio Luís
Department of Pharmaceutical Science, State University of Ponta Grossa, Ponta Grossa, PR, Brazil.
Department of General Biology, State University of Ponta Grossa, Ponta Grossa, PR, Brazil.
J Ethnopharmacol. 2016 May 13;183:29-37. doi: 10.1016/j.jep.2016.02.025. Epub 2016 Feb 22.
Euphorbia umbellata latex (sap) has normally been used in folk medicine in southern Brazil to treat different types of cancers.
To carry out a biomonitored investigation of partitioned latex using in vitro assay, to identify the main mechanisms related with the action of the most active fraction as well as to develop a phytochemical study with this material.
Biological screening was performed with hexane, chloroform, ethyl acetate and methanol fractions from the latex of E. umbellata using MTT, trypan blue, and neutral red assays to determine the cytotoxicity against HRT-18, HeLa and Jurkat cells and flow cytometry, DNA quantification, acridine orange and Hoechst 33342 staining to investigate mechanisms of action for the hexane extract. The phytochemical study of the hexane fraction was performed by chromatographic procedures and the substances were identified by NMR analysis. The isolated terpenes were evaluated using MTT to determine the cytotoxicity against Jurkat cells.
All the fractions presented concentration and time dependent cytotoxicity. The hexane fraction showed the highest cytotoxicity; whereas the Jurkat cell was the lineage with the highest sensitivity (IC50 1.87µg/mL). Fragmentation of DNA and apoptosis are two mechanisms related with the toxicity of hexane fraction. The hexane fraction arrested the cell cycle in the G0/G1 phase, and the selectivity index was 4.30. Phytochemical study of the hexane fraction led to isolation of euphol (main compound) and germanicol acetate. Both substances demonstrated some slight cytotoxic activity against Jurkat cells after 72h; however the activity was minimal compared to vincristine (anticancer standard drug).
The current research proves that the fractions of the latex from E. umbellata have a cytotoxic effect against three different cancer cells lines. The hexane fraction showed high in vitro cytotoxic effects against Jurkat cells demonstrating that the effect may be due to non-polar constituents. The two isolated terpenes (euphol and germanicol acetate) showed poor cytotoxic activity indicating that the anticancer properties of the extract may be caused by other substances present in the hexane fraction.
伞大戟乳胶（汁液）在巴西南部的民间医学中通常用于治疗不同类型的癌症。
使用体外试验对分级乳胶进行生物监测研究，以确定与最具活性部分的作用相关的主要机制，并对该材料进行植物化学研究。
使用MTT、台盼蓝和中性红试验，对伞大戟乳胶的己烷、氯仿、乙酸乙酯和甲醇级分进行生物筛选，以确定对HRT - 18、HeLa和Jurkat细胞的细胞毒性，并通过流式细胞术、DNA定量、吖啶橙和Hoechst 33342染色研究己烷提取物的作用机制。通过色谱程序对己烷级分进行植物化学研究，并通过NMR分析鉴定物质。使用MTT评估分离出的萜类化合物对Jurkat细胞的细胞毒性。
所有级分均呈现浓度和时间依赖性细胞毒性。己烷级分显示出最高的细胞毒性；而Jurkat细胞是最敏感的细胞系（IC50为1.87μg/mL）。DNA片段化和凋亡是与己烷级分毒性相关的两种机制。己烷级分使细胞周期停滞在G0/G1期，选择性指数为4.30。对己烷级分的植物化学研究导致分离出大戟醇（主要化合物）和乙酸germanicol。两种物质在72小时后对Jurkat细胞均表现出一些轻微的细胞毒性活性；然而，与长春新碱（抗癌标准药物）相比，活性最小。
目前的研究证明，伞大戟乳胶的级分对三种不同的癌细胞系具有细胞毒性作用。己烷级分对Jurkat细胞显示出高体外细胞毒性作用，表明该作用可能归因于非极性成分。两种分离出的萜类化合物（大戟醇和乙酸germanicol）显示出较差的细胞毒性活性，表明提取物的抗癌特性可能由己烷级分中存在的其他物质引起。
