泰国部分纯化的黄猄蚁蜂胶对人癌细胞系的体外抗增殖活性。
In vitro antiproliferative activity of partially purified Trigona laeviceps propolis from Thailand on human cancer cell lines.
机构信息
Department of Biology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
出版信息
BMC Complement Altern Med. 2011 May 6;11:37. doi: 10.1186/1472-6882-11-37.
BACKGROUND
Cancers are some of the leading causes of human deaths worldwide and their relative importance continues to increase. Since an increasing proportion of cancer patients are acquiring resistance to traditional chemotherapeutic agents, it is necessary to search for new compounds that provide suitable specific antiproliferative affects that can be developed as anticancer agents. Propolis from the stingless bee, Trigona laeviceps, is one potential interesting source that is widely available and cultivatable (as bee hives) in Thailand.
METHODS
Propolis (90 g) was initially extracted by 95% (v/v) ethanol and then solvent partitioned by sequential extractions of the crude ethanolic extract with 40% (v/v) MeOH, CH₂Cl₂ and hexane. After solvent removal by evaporation, each extract was solvated in DMSO and assayed for antiproliferative activity against five cancer (Chago, KATO-III, SW620, BT474 and Hep-G2) and two normal (HS27 fibroblast and CH-liver) cell lines using the MTT assay. The cell viability (%) and IC₅₀ values were calculated.
RESULTS
The hexane extract provided the highest in vitro antiproliferative activity against the five tested cancer cell lines and the lowest cytotoxicity against the two normal cell lines. Further fractionation of the hexane fraction by quick column chromatography using eight solvents of increasing polarity for elution revealed the two fractions eluted with 30% and 100% (v/v) CH₂Cl₂ in hexane (30DCM and 100DCM, respectively) had a higher anti-proliferative activity. Further fractionation by size exclusion chromatography lead to four fractions for each of 30DCM and 100DCM, with the highest antiproliferative activity on cancer but not normal cell lines being observed in fraction# 3 of 30DCM (IC₅₀ value of 4.09 - 14.7 μg/ml).
CONCLUSIONS
T. laeviceps propolis was found to contain compound(s) with antiproliferative activity in vitro on cancer but not normal cell lines in tissue culture. The more enriched propolis fractions typically revealed a higher antiproliferative activity (lower IC₅₀ value). Overall, propolis from Thailand may have the potential to serve as a template for future anticancer-drug development.
背景
癌症是全球人类死亡的主要原因之一，其重要性持续增加。由于越来越多的癌症患者对传统化疗药物产生耐药性，因此有必要寻找新的化合物，这些化合物能提供合适的特定抗增殖作用，可开发为抗癌药物。来自无刺蜜蜂，T. laeviceps 的蜂胶是一种潜在的有趣来源，在泰国广泛可用且可种植（作为蜂箱）。
方法
首先用 95%（v/v）乙醇提取 90 克蜂胶，然后用 40%（v/v）甲醇、CH₂Cl₂和正己烷依次萃取粗乙醇提取物进行溶剂分配。蒸发除去溶剂后，将每个提取物溶解在 DMSO 中，并通过 MTT 测定法测定其对 5 种癌细胞（Chago、KATO-III、SW620、BT474 和 Hep-G2）和 2 种正常细胞（HS27 成纤维细胞和 CH-肝）的体外增殖活性。计算细胞活力（%）和 IC₅₀ 值。
结果
正己烷提取物对 5 种测试的癌细胞系表现出最高的体外增殖抑制活性，对 2 种正常细胞系的细胞毒性最低。用八种极性递增的溶剂通过快速柱色谱对正己烷馏分进一步分级，发现用 30%和 100%（v/v）CH₂Cl₂洗脱的两个馏分（分别为 30DCM 和 100DCM）具有更高的抗增殖活性。进一步通过尺寸排阻色谱分离得到 30DCM 和 100DCM 的每一个都有四个馏分，在癌细胞系而非正常细胞系中观察到的最高抗增殖活性存在于 30DCM 的第 3 馏分中（IC₅₀ 值为 4.09-14.7μg/ml）。
结论
T. laeviceps 蜂胶中含有在体外对组织培养中的癌细胞具有抗增殖活性的化合物。更富集的蜂胶馏分通常显示出更高的抗增殖活性（更低的 IC₅₀ 值）。总的来说，来自泰国的蜂胶可能有潜力作为未来抗癌药物开发的模板。
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