Chlorogenic acid induced apoptosis and inhibition of proliferation in human acute promyelocytic leukemia HL-60 cells
Chlorogenic acid induced apoptosis and inhibition of proliferation in human acute promyelocytic leukemia HL-60 cells
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DOI：
10.3892/mmr.2013.1652
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发表时间：
2013-10-01
影响因子：
3.4
通讯作者：
Wang, Yong-Tang
中科院分区：
文献类型：
--
作者：
Liu, Ya-Jing;Zhou, Chang-Yang;Wang, Yong-Tang
绿原酸（Chlorogenic acid， CA）在许多植物的叶片中含量很高，具有抗菌、抗炎、抗诱变、抗氧化等生物活性。据报道，它通过诱导慢性髓性白血病（CML）细胞系（包括U937和K562细胞）的凋亡而具有抗肿瘤活性。然而，CA对人类急性早幼粒细胞白血病（APL） HL-60细胞的影响尚不清楚。本研究考察了CA对人APL HL-60细胞的G0/G1周期阻滞和诱导凋亡的作用。1、5、10 μ M CA处理5 d后，用台盼蓝染色法绘制生长曲线，观察CA对HL-60细胞的影响及细胞活力。不同剂量CA处理48 h后，采用Wright’s- giemsa染色、Hoechst 33342染色、碘化丙啶（PI）染色、DNA阶梯分析和流式细胞术观察细胞凋亡诱导和细胞增殖抑制情况。结果表明，72 h时HL-60细胞生长达到平台期，CA处理组HL-60细胞增殖抑制率明显高于对照组。以时间和剂量依赖的方式。然而，Wright’s- giemsa染色、Hoechst 33342染色和PI染色显示，与未用药的细胞相比，CA处理的HL-60细胞的凋亡水平明显升高，形成更多的凋亡小体。通过DNA阶梯分析和流式细胞术发现，CA处理后的细胞有明显的特征DNA阶梯，CA能够将细胞周期阻滞在G0/G1期。CA处理HL-60细胞48 h后，可显著促进HL-60细胞凋亡，并呈剂量依赖性，同时可抑制细胞增殖。结果表明，CA可抑制HL-60细胞增殖，诱导HL-60细胞前期凋亡。因此，10 μ M的浓度可能是治疗人急性早幼粒细胞白血病的最佳剂量。
Chlorogenic acid (CA), is found in high abundance in the leaves of a number of plants and has antibacterial, antiphlogistic, antimutagenic, antioxidant and other biological activities. It reportedly possesses antitumor activity via the induction of apoptosis in chronic myelogenous leukemia (CML) cell lines, including U937 and K562 cells. However, the effects of CA on human acute promyelocytic leukemia (APL) HL-60 cells remains unknown. In the current study, the ability of CA to cause G0/G1 cycle arrest and induce apoptosis in the treatment of human APL HL-60 cells was investigated. Following 5 days treatment with 1, 5 and 10 mu M CA, cell viability and the effects of CA on the growth of HL-60 cells were investigated using a growth curve constructed using trypan blue staining. Induction of apoptosis and inhibition of cell proliferation were estimated using Wright's-Giemsa staining, Hoechst 33342 and propidium iodide (PI) staining, DNA ladder analysis and flow cytometry, following 48 h cell treatment with various doses of CA. The results indicated that the growth of HL-60 cells reached a plateau phase at 72 h and the proliferation inhibition rate of HL-60 cells in CA-treated groups was significantly higher compared with the control, in a time- and dose-dependent manner. However, the level of apoptosis of HL-60 cells treated with CA markedly increased and formed more apoptotic bodies compared with the cells with no drug treatment, according to the Wright's-Giemsa staining, Hoechst 33342 and PI staining, respectively. Using DNA ladder analysis and flow cytometry it was shown that a significant characteristic DNA ladder was observed when treated with CA. CA was capable of arresting cell cycle at G0/G1 phase. Apoptosis of HL-60 cells treated with CA for 48 h was promoted significantly in a dose-dependent manner, as well as the inhibition of proliferation. The observations revealed that CA inhibits proliferation and induces preprophase apoptosis of HL-60 cells. Thus, the concentration of 10 mu M may be the optimal dose for treatment human acute promyelocytic leukemia.