Yu Qinghong, Ma Li, Shen Yiping, Zhai Wo, Zhou Yuhong
Department of Hematology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310006, China.
Zhonghua Xue Ye Xue Za Zhi. 2015 Nov;36(11):937-41. doi: 10.3760/cma.j.issn.0253-2727.2015.10.010.
To investigate the effects of angular pyranocoumarin (±) -4'-O- acetyl-3'-Oangeloyl- cis- khellactone (APC) extracted from peucedanum praeruptoruon on the proliferation and apoptosis of U266 cells, and to explore its related mechanism.
APC was extracted by petroleum ether technique, and its purity was tested by high performance liquid chromatography, and its chemical structure was identified by magnetic resonance spectroscopy. U266 cells were treated with APC in various concentrations (0, 10, 20, 30, 40 μg/ml）for different durations（24 and 48 h). The inhibitive effect of APC on cell growth was detected by CCK-8 method. After U266 cells were incubated with APC（0, 10, 20, 30, 40 μg/ml）for 24 h, the apoptosis of cells were observed by flow cytometry stained with Annexin Ⅴ/PI and Hochest33342; the expression levels of caspase-3, 8, ERK, p-ERK, AKT and p-AKT protein were assayed by Western blot; the expression of hTERT mRNA was measured by RT-PCR.
The purity of APC identified by magnetic resonance imaging was 98.8%. The proliferation of U266 cells was inhibited, and the apoptosis was induced in a time- and/or dose- dependent manner after treatment with APC. APC could upregulate the caspase- 8, 3 protein expression and downregulate the p- ERK, p-AKT protein expression along with the increase of APC dose. APC also could downregulate the hTERT mRNA expression.
Angular pyranocoumarin APC could inhibit the proliferation and induce the apoptosis of U266 cells. The probable mechanism might be achieved by upregulating caspase-8, 3 protein expression and downregulating p-ERK, P-AKT protein and the hTERT mRNA expression.
研究从白花前胡中提取的角型吡喃香豆素（±）-4'-O-乙酰基-3'-O-当归酰基-顺式凯刺内酯（APC）对U266细胞增殖和凋亡的影响，并探讨其相关机制。
采用石油醚法提取APC，用高效液相色谱法检测其纯度，用磁共振波谱法鉴定其化学结构。将U266细胞用不同浓度（0、10、20、30、40μg/ml）的APC处理不同时间（24和48小时）。用CCK-8法检测APC对细胞生长的抑制作用。将U266细胞与APC（0、10、20、30、40μg/ml）孵育24小时后，用AnnexinⅤ/PI和Hochest33342染色，通过流式细胞术观察细胞凋亡情况；用蛋白质免疫印迹法检测caspase-3、8、ERK、p-ERK、AKT和p-AKT蛋白的表达水平；用逆转录聚合酶链反应（RT-PCR）检测hTERT mRNA的表达。
磁共振成像鉴定的APC纯度为98.8%。APC处理后，U266细胞的增殖受到抑制，凋亡呈时间和/或剂量依赖性诱导。随着APC剂量的增加，APC可上调caspase-8、3蛋白表达，下调p-ERK、p-AKT蛋白表达。APC还可下调hTERT mRNA表达。
角型吡喃香豆素APC可抑制U266细胞的增殖并诱导其凋亡。其可能的机制可能是通过上调caspase-8、3蛋白表达，下调p-ERK、P-AKT蛋白和hTERT mRNA表达来实现的。
