<p>Notopterol-induced apoptosis and differentiation in human acute myeloid leukemia HL-60 cells</p>

Abstract

Purpose: This study aims to observe the effects of notopterol on the apoptosis and differentiation of HL-60 cells and to explore the underlying molecular mechanisms. Methods: Cell viability was assessed using sulforhodamine B assay. Cell proliferation was determined by the trypan blue dye exclusion test. Colony-forming units were assayed in methylcellulose. Apoptosis assays were carried out by annexin V-fluorescein isothiocyanate(FITC)/propidium iodide (PI) double staining, Hoechst 33342 staining, mitochondrial membrane potential, and Western blot. Wright–Giemsa staining, nitroblue tetrazolium (NBT) reduction assay, CD11b and CD14 and Western blot were detected for induction of differentiation. In addition, cell-cycle phase distribution was analyzed by flow cytometry and Western blot. The combination therapy of notopterol and all-trans retinoic acid (ATRA) on HL-60 cells was examined. Results: Notopterol obviously inhibited the growth of HL-60 cells with an IC 50 value of 40.32 μM and remarkably reduced the number of colonies by 10, 20, and 40 µM. In addtion, notopterol induced the percentage of apoptotic HL-60 cells, reduced the mitochondrial membrane potential, decreased the protein expresstion of Bcl-2 and Mcl-1, and increased the expression of Bax, cleavage of caspase 9, caspase 3, and PARP. As for cell differentiation, notopterol clearly induced chromatin condensation; increased the nucleocytoplasmic ratio, nitroblue tetrazolium-positive cells, expression of CD14 and CD11b, and protein expression of c-Jun and Jun B, and decreased c-myc. Furthermore, notopterol induced the G0/G1 cell-cycle arrest as determined using flow cytometry, which may be related to the regulation of cell-cycle-related proteins p53, CDK2, CDK4, Cyclin D1, Cyclin E, and survivin. The combined use of notopterol and ATRA did not enhance the apoptotic effect as evidenced by cell viability test and Hoechst 33342. However, the combination of notopterol and ATRA enhanced the effect of inducing differentiation when compared with using either notopterol or ATRA alone, which can be evidenced by the increased nucleocytoplasmic ratio, NBT positive cells, and expression of CD14. Conclusion: This is the first time it has been demonstrated that notopterol could induce apoptosis, differentiation, and G0/G1 arrest in human AML HL-60 cells, suggesting that notopterol has potential therapeutic effects on AML. The combination application of notopterol (20 and 40 μM) and ATRA (2 μM) could augment differentiation of HL-60 cells.

Isobavachalcone inhibits acute myeloid leukemia: Potential role for ROS‐dependent mitochondrial apoptosis and differentiation

“…The proteinbound dye was dissolved in 10 mM Tris base solution for OD determination at a wavelength of 490 nm using a multiwell spectrophotometer microplate reader (Biotek, Winooski, VT). Cell viability was presented as a percentage of that of untreated cells (Huang et al, 2019).…”

“…Morphological assessment of HL-60 and Kasumi-1 cells or peripheral blood obtained from NOD/SCID mice was performed using Wright-Giemsa (Leagene, Beijing, China) staining according to the manufacturer's protocol on slides prepared by cytospin. The morphology of cells was examined under a light microscope (Huang et al, 2019).…”

Imidazo[1,2-b]pyrazole-7-Carboxamide Derivative Induces Differentiation-Coupled Apoptosis of Immature Myeloid Cells Such as Acute Myeloid Leukemia and Myeloid-Derived Suppressor Cells

“…In order to study the underlying mechanism of action of our recently developed drug candidate, DU325 with anti-leukemic effect [21], the prototype model of human acute myeloblastic leukemia, the HL-60 cells, were treated and assayed. HL-60 cells have been widely used earlier to test differentiation induced chemotherapy [36][37][38]. We have previously observed the asynchronous response of HL-60 cells in terms of morphology treated with DU325 and decided to obtain single cell protein expression data in a subpopulation of cells (bright) by using flow cytometry.…”

Role of 2′-Hydroxycinnamaldehyde In The Induction of Apoptosis Via A Reactive Oxygen Species-Dependent JNK Pathway in Human Promyelocytic HL-60 Leukemia Cells

“…Leukemia is a major hematological malignancy that causes mortality and morbidity in different age groups [4]. It is de ned as the abnormal proliferation, clonality, and differentiation of immature hematopoietic cells in the bone marrow [5]. Leukemia can be classi ed into four common types: acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), and chronic myelogenous leukemia (CML).…”

“…Among these types of leukemia, AML is characterized by the aggressive growth of hematopoietic precursor cells that interfere with the production of normal hematopoietic cells in the bone marrow [6]. Although re nement of supportive treatment has improved the outlook of patients with AML in the past 30 years, more than half of young adults and around 90% of older patients still die from AML [5]. The majority of AML cells express varying amounts of the transmembrane surface glycoprotein CD33 (observed in approximately > 80% of patients with AML).…”