Abstract
Tetrahydroisoquinolines are known to have various biological effects, including antitumor activity. This study investigated the effect of 1-chloromethyl-6, 7-dimethoxy-3, 4-dihydro-1H-isoquinoline-2-sulfonic acid amide (CDST), a newly synthesized anticancer agent, on cellular differentiation and proliferation in HL-60 cells. Differentiation and proliferation of HL-60 cells were determined through expression of CD11b and CD14 surface antigens using flow cytometry and nitroblue tetrazolium (NBT) assay, and through analysis of cell cycle using propidium iodide staining, western blot analysis and immunoprecipitation, respectively. CDST induced the differentiation of HL-60, as shown by increased expression of differentiation surface antigen CD11b (but no significant change in CD14 expression) and increased NBT-reducing functional activity. DNA flow cytometry analysis indicated that CDST markedly induced a G0/G1 phase arrest of HL-60 cells. Subsequently, we examined the expre-ssion of G0/G1 phase cell cycle-related proteins, including cyclin-dependent kinases (CDKs), cyclins and cyclin dependent kinase inhibitors (CKIs), during the differentiation of HL-60. The levels of CDK2, CDK6, cyclin E and cyclin A were decreased, whereas steady-state levels of CDK4 and cyclin D1 were unaffected. The expression of the p27Kip1 was markedly increased by CDST, but not p21WAF1/Cip1. Moreover, CDST markedly enhanced the binding of p27Kip1 with CDK2 and CDK6, resulting in the reduced activity of both kinases. Taken together, these results demonstrate that CDST is capable of inducing cellular differentiation and growth inhibition through p27Kip1 protein-related G0/G1 phase arrest in HL-60 cells.
Highlights
Treatment of the promyelocytic leukemia cell lines, including HL-60 cells, with anti-leukemic agents induces differentiation into two cell types of the myeloid lineage: monocyte/macrophage-like cells [1,2] and granulocytelike cells [3]. 12-O-Tetradecanoyl-phorbol-13-acetate and all-trans-retinoic acid (ATRA) are well-known inducers that can stimulate differentiation of HL-60 cells into monocyte/macrophage-like and granulocyte-like phenoltypes, respectively [1,2,4]
We examined whether CDST could induce differentiation of HL-60 cells
HL-60 cells were treated with CDST or ATRA, and the numbers of differentiated cells were determined by measuring nitroblue tetrazolium (NBT)-reducing activity
Summary
Treatment of the promyelocytic leukemia cell lines, including HL-60 cells, with anti-leukemic agents induces differentiation into two cell types of the myeloid lineage: monocyte/macrophage-like cells [1,2] and granulocytelike cells [3]. 12-O-Tetradecanoyl-phorbol-13-acetate and all-trans-retinoic acid (ATRA) are well-known inducers that can stimulate differentiation of HL-60 cells into monocyte/macrophage-like and granulocyte-like phenoltypes, respectively [1,2,4]. 12-O-Tetradecanoyl-phorbol-13-acetate and all-trans-retinoic acid (ATRA) are well-known inducers that can stimulate differentiation of HL-60 cells into monocyte/macrophage-like and granulocyte-like phenoltypes, respectively [1,2,4]. After exposure to these agents, HL-60 cells tend to stop cellular proliferating and express the phenotypical and physiological functions specific to mature cells. Ju et al / Health 5 (2013) 1-7 gression in mammalian cells is regulated by a family of enzymes known as cyclin-dependent kinases (CDKs) whose activity is dependent on the binding to specific regulatory subunits called cyclins [5,8]. The mechanisms for regulation of cell cycle have a fundamental role in the control of cell proliferation and may be involved in differentiation
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