Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur 50603, Malaysia.
Department of Chemistry, Faculty of Science, University of Malaya, Kuala Lumpur 50603, Malaysia.
Molecules. 2018 Oct 23;23(11):2733. doi: 10.3390/molecules23112733.
Pinnatane A from the bark of was investigated for its anti-cancer properties by analyzing the cytotoxic activities and cell cycle arrest mechanism induced in two different liver cancer cell lines.
A 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used to analyze the pinnatane A selectivity in inducing cell death in cancer and normal cells. Various biological assays were carried out to analyze the anti-cancer properties of pinnatane A, such as a live/dead assay for cell death microscopic visualization, cell cycle analysis using propidium iodide (PI) to identify the cell cycle arrest phase, annexin V-fluorescein isothiocyanate (annexin V-FITC)/PI flow cytometry assay to measure percentage of cell populations at different stages of apoptosis and necrosis, and DNA fragmentation assay to verify the late stage of apoptosis.
The MTT assay identified pinnatane A prominent dose- and time-dependent cytotoxicity effects in Hep3B and HepG2 cells, with minimal effect on normal cells. The live/dead assay showed significant cell death, while cell cycle analysis showed arrest at the G₀/G₁ phase in both cell lines. Annexin V-FITC/PI flow cytometry and DNA fragmentation assays identified apoptotic cell death in Hep3B and necrotic cell death in HepG2 cell lines.
Pinnatane A has the potential for further development as a chemotherapeutic agent prominently against human liver cells.
从白皮肉桂的树皮中分离得到的 pinnatane A,通过分析其在两种不同肝癌细胞系中诱导的细胞毒性活性和细胞周期阻滞机制,研究其抗癌特性。
采用 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2H-四唑溴盐(MTT)法分析 pinnatane A 在诱导癌细胞和正常细胞死亡方面的选择性。进行了各种生物测定来分析 pinnatane A 的抗癌特性,例如细胞死亡的活/死检测用于细胞死亡的微观可视化,碘化丙啶(PI)用于细胞周期分析以鉴定细胞周期阻滞相,膜联蛋白 V-异硫氰酸荧光素(annexin V-FITC)/PI 流式细胞术检测用于测量不同凋亡和坏死阶段的细胞群体百分比,以及 DNA 片段化检测用于验证晚期凋亡。
MTT 测定法确定了 pinnatane A 在 Hep3B 和 HepG2 细胞中具有明显的剂量和时间依赖性细胞毒性作用,对正常细胞的影响最小。活/死检测显示出明显的细胞死亡,而细胞周期分析显示在两种细胞系中均在 G₀/G₁ 期停滞。Annexin V-FITC/PI 流式细胞术和 DNA 片段化检测鉴定出 Hep3B 细胞的凋亡性细胞死亡和 HepG2 细胞的坏死性细胞死亡。
Pinnatane A 具有进一步开发为化学治疗剂的潜力,特别是针对人类肝细胞。