Department of Biomedical Sciences, Faculty of Medicine, Prince of Songkla University, Hat Yai, Songkhla 90110, Thailand.
The Cancer Molecular Biology Excellence Research Laboratory, Prince of Songkla University, Hat Yai, Songkhla 90110, Thailand.
Nutrients. 2015 Apr 10;7(4):2707-18. doi: 10.3390/nu7042707.
This study aimed to evaluate the cytotoxicity of a crude extract of Piper cubeba against normal and breast cancer cell lines. To prepare the extract, P. cubeba seeds were ground, soaked in methanol and dichloromethane and isolated by column chromatography. Fractions were tested for cytotoxicity effects on normal fibroblast (L929), normal breast (MCF-12A) and breast cancer cell lines (MCF-7, MDA-MB-468 and MDA-MB-231). The most effective fraction was selected for DNA fragmentation assay to detect apoptotic activity. The results showed that the methanolic crude extract had a higher cytotoxic activity against MDA-MB-468 and MCF-7 than a dichloromethane crude extract. Then, the methanolic crude extract was separated into six fractions, designated A to F. Fraction C was highly active against breast cancer cell lines with an IC50 value less than 4 μg/mL. Therefore, Fraction C was further separated into seven fractions, CA to CG. The 1H-NMR profile showed that Fraction CE was long chain hydrocarbons. Moreover, Fraction CE demonstrated the highest activity against MCF-7 cells with an IC50 value of 2.69 ± 0.09 μg/mL and lower cytotoxicity against normal fibroblast L929 cells with an IC50 value of 4.17 ± 0.77 μg/mL. Finally, DNA fragmentation with a ladder pattern characteristic of apoptosis was observed in MCF-7, MDA-MB-468, MDA-MB-231 and L929 cells, but not in MCF-12A cells.
本研究旨在评估胡椒（Piper cubeba）粗提取物对正常和乳腺癌细胞系的细胞毒性。为了制备提取物，将胡椒种子研磨，用甲醇和二氯甲烷浸泡，然后通过柱层析分离。测试各馏分对正常成纤维细胞（L929）、正常乳腺（MCF-12A）和乳腺癌细胞系（MCF-7、MDA-MB-468 和 MDA-MB-231）的细胞毒性作用。选择最有效的馏分进行 DNA 片段化分析以检测细胞凋亡活性。结果表明，与二氯甲烷粗提取物相比，甲醇粗提取物对 MDA-MB-468 和 MCF-7 的细胞毒性更高。然后，甲醇粗提取物被分离成六个馏分，分别标记为 A 到 F。馏分 C 对乳腺癌细胞系具有高度活性，IC50 值小于 4 μg/mL。因此，馏分 C 进一步分离成七个馏分，CA 到 CG。1H-NMR 图谱显示馏分 CE 是长链烃。此外，馏分 CE 对 MCF-7 细胞表现出最高的活性，IC50 值为 2.69 ± 0.09 μg/mL，对正常成纤维细胞 L929 细胞的细胞毒性较低，IC50 值为 4.17 ± 0.77 μg/mL。最后，在 MCF-7、MDA-MB-468、MDA-MB-231 和 L929 细胞中观察到具有凋亡特征的梯状 DNA 片段化，但在 MCF-12A 细胞中未观察到。
