8-氯腺苷对未分化HL-60细胞增殖的影响

摘要/Abstract

摘要： 目的 8-氯腺苷对HL-60细胞增殖抑制作用及其机制的探讨. 方法采用MTT和SRB方法检测了8-氯腺苷对不同种类人肿瘤细胞增殖的作用; 通过扫描电镜和透射电镜观察了HL-60细胞表面和核的变化; 利用NBT还原和酸性磷酸酶分析诱导HL-60细胞分化; 用流式细胞仪分析了8-氯腺苷对HL-60细胞周期的影响. 结果 8-氯腺苷对七种人肿瘤细胞的增殖有抑制作用, 其IC50值为KB(0.05μmol·L^-1)<HL-60(0.25 μmol·L^-1)<Bel-7402(0.56μmol·L^-1)<MCF(0.65μmol·L^-1)<HCT(0.79μmol·L^-1)<Hela(0.89μmol·L^-1)<BGC-823(1.49μmol·L^-1)<PG(2.50μmol·L^-1).电镜结果显示, 8-氯腺苷(0.5μmol·L^-1)与HL-60细胞作用72 h, HL-60细胞表面微绒毛变短, 细胞核型发生了改变, 如马蹄、肾和分叶型.同时8-氯腺苷可促进HL-60细胞对NBT的还原能力及增加细胞中酸性磷酸酶的活性, 其作用具有时间和浓度依赖性, 说明8-氯腺苷可诱导HL-60细胞的分化. 细胞周期分析证实, 8-氯腺苷使HL-60细胞G1期增多, S期细胞减少. 结论 8-氯腺苷可以诱导HL-60细胞的分化, 并阻止HL-60细胞停止在G1期,从而导致HL-60细胞增殖抑制.

关键词: 8-氯腺苷, HL-60细胞, 诱导分化, 细胞周期

Abstract: Aim To study the effect of 8-chloroadenosine (8-CA)on undifferentiatied HL-60 cell line. Methods The IC50 of cancer cell proliferation was determined using a microculture plate reader at 570 nm (MTT) and 540 nm (SRB). Morphology of HL-60 cells was observed under a scanning electron microscope and a transmission electron microscope. The differentiation of HL-60 cells was examined by nitro blue tetrazolium reduction (NBT) and acid phosphatase assay. The cy-cle of HL-60 cells was analyzed by flow cytometry. Results 8-CA inhibited proliferation of eight human cancer cell lines.The IC50 ranked in the following order: KB (0.05μmol·L^-1)<HL-60(0.25 μmol·L^-1)<Bel-7402(0.56μmol·L^-1)<MCF(0.65μmol·L^-1)<HCT(0.79μmol·L^-1)<Hela(0.89μmol·L^-1)<BGC-823(1.49μmol·L^-1)<PG(2.50μmol·L^-1). The scanning and transmission electron microscopy showed that the microvilli of HL-60 cell surface shortened, and the shape of HL-60 cells nuclei changed to kidney-shaped, horse shoe-shaped and bilob ated after treatment with 8-CA. Meanwhile, 8-CA promoted NBT reduction and increased activity of acid phosphatase in HL-60 cells in a time and concentration-dependent manner. Flow cytometry analysis indicated that 8-CA induced an appreciable increase of the cell population in G1 phase with a marked reduction in S phase. Conclusion 8-CA can induce differentiation of HL-60 cells and block the cells at G1 phase, thus inhibiting proliferation of HL-60 cells.

Key words: 8-chloroadenosine, 8-chloroadenosine, HL-60 cell line, HL-60 cell line, differentiation, differentiation, cell cycle, cell cycle

中图分类号:

R965.2

R979.1

Supporting: ^*Corresponding author. Tel.: 010-82801700

崔景荣, 惠峪, 相幼青, 张礼和^*. 8-氯腺苷对未分化HL-60细胞增殖的影响[J]. .

CUI Jing-rong, HUI Yu, XIANG You-qing, ZHANG Li-he^*. Effect of 8-Chloroadenosine on Undifferentiatied HL-60 Cell Line[J]. .

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