College of Pharmacy and Bioengineering, Chongqing University of Technology, Chongqing 400054, P.R. China.
Mol Med Rep. 2013 Oct;8(4):1106-10. doi: 10.3892/mmr.2013.1652. Epub 2013 Aug 27.
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 µM 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 µM may be the optimal dose for treatment human acute promyelocytic leukemia.
绿原酸（CA）在许多植物的叶子中含量丰富，具有抗菌、抗炎、抗突变、抗氧化等多种生物学活性。据报道，CA 通过诱导慢性髓系白血病（CML）细胞系，包括 U937 和 K562 细胞的凋亡，具有抗肿瘤活性。然而，CA 对人急性早幼粒细胞白血病（APL）HL-60 细胞的影响尚不清楚。在本研究中，研究了 CA 诱导 G0/G1 期细胞周期阻滞和诱导人 APL HL-60 细胞凋亡的能力。用台盼蓝染色法绘制生长曲线，研究了 1、5 和 10μM CA 处理 5 天后对细胞活力和 CA 对 HL-60 细胞生长的影响。用 Wright-Giemsa 染色、Hoechst 33342 和碘化丙啶（PI）染色、DNA 梯状分析和流式细胞术分别检测 CA 处理 48 小时后细胞凋亡和增殖抑制的情况。结果表明，HL-60 细胞的生长在 72 小时达到平台期，CA 处理组 HL-60 细胞的增殖抑制率明显高于对照组，呈时间和剂量依赖性。然而，与无药物处理的细胞相比，CA 处理的 HL-60 细胞的凋亡水平明显增加，形成更多的凋亡小体，根据 Wright-Giemsa 染色、Hoechst 33342 和 PI 染色分别观察到。用 DNA 梯状分析和流式细胞术观察到 CA 处理后出现明显的特征性 DNA 梯状。CA 能够将细胞周期阻滞在 G0/G1 期。CA 处理 48 小时后，HL-60 细胞的凋亡呈剂量依赖性显著增加，同时抑制增殖。观察结果表明，CA 抑制 HL-60 细胞的增殖，并诱导其前期凋亡。因此，10μM 的浓度可能是治疗人急性早幼粒细胞白血病的最佳剂量。
