Mohammadi Saeed, Zahedpanah Mahdi, Ghaffari Seyed Hamidollah, Shaiegan Mojgan, Nikbakht Mohsen, Nikugoftar Mahin
Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Department of Medical laboratory sciences, Faculty of Allied Medicine, Qazvin University of Medical Sciences, Qazvin, Iran.
Life Sci. 2017 Nov 15;189:89-95. doi: 10.1016/j.lfs.2017.09.019. Epub 2017 Sep 19.
To determine if parthenolide (PTL) is cytotoxic for leukemia-like KG1a cells and if it involves in certain molecular-mediated resistance, especially osteopontin (OPN).
PTL/daunorubicin (DNR)-treated KG1a cells were examined for viability using MTT and colony-formation assay, and stained for apoptosis using AV/PI. The gene and protein expression were evaluated by qReal-time PCR and Western blotting analysis, respectively. OPN gene was inhibited by OPN siRNA. The cells were stained for various fractions using PE anti-CD34, FITC anti-CD38 and PerCP anti-CD123.
Cell viability and proliferation assay exhibited KG1a cells are relatively refractory to used concentrations of PTL. OPN mRNA and protein levels increased in response to PTL. Suppression of OPN with siRNA increased the cytotoxic effects of PTL on KG1a cells. PTL treatment and OPN siRNA suppression in KG1a cells resulted in a decrease of mRNA expression of AKT, mTOR, β-catenin, and Phosphatase and tensin homolog (PTEN). The sub-population cells of CD34 and CD123 from KG1a cells are enriched by PTL treatment.
Parthenolide in spite of the reduction in gene expression of AKT, mTOR or beta-catenin, stimulates the OPN expression in KG1a cells. The OPN expression pattern in KG1a cells could be compatible with CD34/CD123 subtype enrichment by PTL which in turn implies OPN's unique role in resistance of cell populations characterized by CD34/CD123 phenotype.
确定小白菊内酯（PTL）对白血病样KG1a细胞是否具有细胞毒性，以及它是否涉及某些分子介导的耐药性，特别是骨桥蛋白（OPN）。
使用MTT和集落形成试验检测经PTL/柔红霉素（DNR）处理的KG1a细胞的活力，并用AV/PI染色检测细胞凋亡。分别通过q实时PCR和蛋白质印迹分析评估基因和蛋白质表达。OPN基因被OPN siRNA抑制。使用PE抗CD34、FITC抗CD38和PerCP抗CD123对细胞进行各种组分染色。
细胞活力和增殖试验表明，KG1a细胞对所用浓度的PTL相对耐药。PTL处理后，OPN mRNA和蛋白质水平升高。用siRNA抑制OPN可增加PTL对KG1a细胞的细胞毒性作用。PTL处理和OPN siRNA抑制导致KG1a细胞中AKT、mTOR、β-连环蛋白和磷酸酶及张力蛋白同源物（PTEN）的mRNA表达降低。PTL处理可富集KG1a细胞中CD34和CD123的亚群细胞。
尽管PTL可降低AKT、mTOR或β-连环蛋白的基因表达，但它仍能刺激KG1a细胞中OPN的表达。KG1a细胞中OPN的表达模式可能与PTL富集CD34/CD123亚型有关，这反过来意味着OPN在以CD34/CD123表型为特征细胞群体的耐药性中具有独特作用。
