19-去甲-1α,25-二羟基维生素D₂（帕立骨化醇）：对人白血病细胞系克隆增殖、分化及凋亡的影响

19-nor-1alpha,25-dihydroxyvitamin D(2) (paricalcitol): effects on clonal proliferation, differentiation, and apoptosis in human leukemic cell lines.

作者信息

Molnár István, Kute Timothy, Willingham Mark C, Powell Bayard L, Dodge William H, Schwartz Gary G

机构信息

Department of Internal Medicine, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA.

出版信息

J Cancer Res Clin Oncol. 2003 Jan;129(1):35-42. doi: 10.1007/s00432-002-0405-7. Epub 2003 Feb 12.

DOI:10.1007/s00432-002-0405-7

PMID:12618899

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12161907/

Abstract

PURPOSE

19-Nor-1alpha,25-dihydroxyvitamin D(2) (paricalcitol) is an analogue of 1,25(OH)(2)D(3) with reduced calcemic effects that is approved for the suppression of parathyroid hormone in chronic renal failure. Paricalcitol has recently been reported to have anticancer activity in prostate cancer. In order to explore paricalcitol as a potential agent against leukemia, we tested its effects on HL-60 and U937 leukemia cell lines.

METHODS

We studied cellular differentiation via expression of CD11b and CD14 surface antigens using flow cytometry, and via the nitroblue tetrazolium (NBT) assay. Cell cycle was analyzed using propidium iodide staining. Apoptosis was assessed with the annexin V assay. Cellular proliferation was determined via colony inhibition on semisolid medium.

RESULTS

Paricalcitol induced the maturation of HL-60 and U937 cells, as shown by increased expression of CD11b differentiation surface antigen. CD14 showed increased expression in HL-60 but not in U937 cells. After exposure to paricalcitol at 10(-8) M for 72 h, the ability of HL-60 cells to reduce NBT was markedly increased. Conversely, U937 cells were unchanged. Paricalcitol inhibited colony formation of both HL-60 and U937 cell lines in semisolid medium after a 10-day incubation (estimated IC(50) of 3x10(-8) M in HL-60 cells and 4x10(-8) M in U937 cells). Paricalcitol at 10(-8) M and 10(-7) M caused a significant dose- and time-dependent increase of apoptosis in HL-60 cells ( P<0.05). In both HL-60 and U937 cells, exposure to 10(-7) M paricalcitol for 72 h increased the number of cells in G(0)/G(1) phase, and decreased the number of cells in S phase.

CONCLUSIONS

Paricalcitol inhibits colony formation, induces maturation and causes cell cycle arrest in HL-60 and U937 cells. Additionally, paricalcitol induces apoptosis in HL-60 cells. These findings support the further evaluation of paricalcitol as an antileukemia agent.

摘要

目的

19-去甲-1α,25-二羟基维生素D₂（帕立骨化醇）是1,25(OH)₂D₃的类似物，其血钙升高作用减弱，已被批准用于抑制慢性肾衰竭患者的甲状旁腺激素。最近有报道称帕立骨化醇在前列腺癌中具有抗癌活性。为了探索帕立骨化醇作为一种潜在的抗白血病药物，我们测试了其对HL-60和U937白血病细胞系的作用。

方法

我们通过流式细胞术检测CD11b和CD14表面抗原的表达以及通过硝基蓝四氮唑（NBT）试验研究细胞分化。使用碘化丙啶染色分析细胞周期。用膜联蛋白V试验评估细胞凋亡。通过半固体培养基上的集落抑制来测定细胞增殖。

结果

帕立骨化醇诱导HL-60和U937细胞成熟，表现为CD11b分化表面抗原表达增加。CD14在HL-60细胞中表达增加，但在U937细胞中未增加。在10⁻⁸M帕立骨化醇作用72小时后，HL-60细胞还原NBT的能力明显增强。相反，U937细胞没有变化。在半固体培养基中孵育10天后，帕立骨化醇抑制HL-60和U937细胞系集落形成（HL-60细胞的估计IC₅₀为3×10⁻⁸M，U937细胞为4×10⁻⁸M）。10⁻⁸M和10⁻⁷M的帕立骨化醇导致HL-60细胞凋亡显著增加，呈剂量和时间依赖性（P<0.05）。在HL-60和U937细胞中，暴露于10⁻⁷M帕立骨化醇72小时会增加G₀/G₁期细胞数量，并减少S期细胞数量。