Zahedpanah Mahdi, Shaiegan Mojgan, Ghaffari Seyed Hamidollah, Nikbakht Mohsen, Nikugoftar Mahin, Mohammadi Saeed
Blood Transfusion Research center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Rep Biochem Mol Biol. 2016 Apr;4(2):82-8.
Interfering with cell proliferation and survival is a critical role for antineoplastic drugs leading to cell death through induction of apoptosis. Alternative treatments with herbal extracts offer insights into acute myeloid leukemia (AML) therapy. Parthenolide (PTL), an extract from feverfew, induces apoptosis in primary human leukemia stem cells (LSCs) and bulk leukemic cell populations. Osteopontin (OPN) preserves cell viability in response to anticancer agents and its receptors could be utilized for therapeutic targeting of cancer cells.
U937 cells were cultured in RPMI 1640 with concentrations of 2, 4, 6, 8, and 10 µM PTL for 20-24 hours for MTT assays. Apoptosis assays were performed with Annexin V-Alexa Fluor-488/PI as Annexin V+/PI- and Annexin V+/PI+ to measure early and late apoptosis, respectively. Quantitative real-time PCR was used to measure OPN gene expression using the 2(-ΔΔCt) method. The PTL-treated cells were stained with FITC-CD38 antibody for flow cytometry analyses. Data were compared using one-way analysis of variance (ANOVA) by SPSS 19.
Parthenolide inhibited growth of U937 cells with IC25 and IC50 values of 4 and 5.8 µM, respectively. Death induction with PTL was apoptotic. Flow cytometry showed a significant decrease in the percentage of CD38+ U937 cells in response to PTL. Osteopontin gene expression decreased in response to PTL.
PTL induced apoptosis and reduced OPN gene expression in U937 cells.
干扰细胞增殖和存活是抗肿瘤药物通过诱导凋亡导致细胞死亡的关键作用。草药提取物的替代疗法为急性髓系白血病（AML）治疗提供了新见解。小白菊内酯（PTL）是一种从小白菊中提取的物质，可诱导原代人白血病干细胞（LSCs）和大量白血病细胞群体凋亡。骨桥蛋白（OPN）可维持细胞对抗癌药物的活力，其受体可用于癌细胞的治疗靶向。
将U937细胞培养于含有浓度为2、4、6、8和10 μM PTL的RPMI 1640培养基中20 - 24小时，用于MTT试验。采用Annexin V - Alexa Fluor - 488/PI进行凋亡试验，分别以Annexin V+/PI-和Annexin V+/PI+测量早期和晚期凋亡。使用2(-ΔΔCt)方法通过定量实时PCR测量OPN基因表达。用FITC - CD38抗体对PTL处理的细胞进行染色，用于流式细胞术分析。数据采用SPSS 19进行单因素方差分析（ANOVA）比较。
小白菊内酯抑制U937细胞生长，IC25和IC50值分别为4和5.8 μM。PTL诱导的死亡为凋亡性。流式细胞术显示，PTL处理后CD38+ U937细胞百分比显著降低。骨桥蛋白基因表达随PTL处理而降低。
PTL诱导U937细胞凋亡并降低OPN基因表达。
