Adan Gökbulut Aysun, Yaşar Mustafa, Baran Yusuf
İzmir Institute of Technology Faculty of Science, Department of Molecular Biology and Genetics, İzmir, Turkey Phone: +90 232 750 73 15 E-mail:
Turk J Haematol. 2015 Jun;32(2):118-26. doi: 10.4274/tjh.2013.0381.
The aims of this study were to examine the cytotoxic and apoptotic effects of KL-21, a novel plant product (produced by Naturin Natural Products, İzmir, Turkey), on 232B4 chronic lymphocytic leukemia (CLL) cells and to determine the cytotoxic effects on healthy BEAS-2B human bronchial epithelial cells.
The cytotoxic effect of KL-21 was determined by MTT cell proliferation assay. Changes in caspase-3 enzyme activity were measured using the caspase-3 colorimetric assay. Changes in mitochondrial membrane potential were determined using the JC-1 dye-based method. Annexin V-FITC/PI double staining was performed to measure the apoptotic cell population. Effects of KL-21 on cell cycle profiles of CLL cells were investigated by flow cytometry.
We detected time- and concentration-dependent increases in the cytotoxic effect of KL-21 on 232B4 CLL cells. However, we also showed that, especially at higher concentrations, KL-21 was less cytotoxic towards BEAS-2B healthy cells than towards CLL cells. Annexin-V/PI double staining results showed that the apoptotic cell population increased in 232B4 cells. Increasing concentrations of KL-21 increased caspase-3 enzyme activity and induced loss of mitochondrial membrane potential. KL-21 administration resulted in small increases in the percentage of the cells in the G0/G1 phase while it decreased the S phase cell population up to 1 mg/mL. At the highest concentration, most of the cells accumulated in the G0/G1 phase.
KL-21 has a growth-inhibitory effect on 232B4 CLL cells. KL-21 causes apoptosis and cell cycle arrest at G0/G1.
本研究旨在检测新型植物产品KL - 21（由土耳其伊兹密尔的Naturin天然产品公司生产）对232B4慢性淋巴细胞白血病（CLL）细胞的细胞毒性和凋亡作用，并确定其对健康的BEAS - 2B人支气管上皮细胞的细胞毒性作用。
采用MTT细胞增殖试验测定KL - 21的细胞毒性作用。使用比色法测定caspase - 3酶活性的变化。采用基于JC - 1染料的方法测定线粒体膜电位的变化。进行Annexin V - FITC/PI双重染色以检测凋亡细胞群体。通过流式细胞术研究KL - 21对CLL细胞周期分布的影响。
我们检测到KL - 21对232B4 CLL细胞的细胞毒性作用呈时间和浓度依赖性增加。然而，我们也表明，尤其是在较高浓度下，KL - 21对BEAS - 2B健康细胞的细胞毒性低于对CLL细胞的细胞毒性。Annexin - V/PI双重染色结果显示232B4细胞中的凋亡细胞群体增加。KL - 21浓度的增加会增加caspase - 3酶活性并诱导线粒体膜电位丧失。给予KL - 21导致G0/G1期细胞百分比略有增加，而在高达1 mg/mL的浓度下，S期细胞群体减少。在最高浓度下，大多数细胞积聚在G0/G1期。
KL - 21对232B4 CLL细胞具有生长抑制作用。KL - 21可导致细胞凋亡并使细胞周期停滞在G0/G1期。
