Cui Xiangzhen, Zhou Shuai, Xu Huanli, Zhao Ting, Liu Aihua, Guo Xueping, Wang Fengshan
Institute of Biochemical and Biotechnological Drugs, School of Pharmaceutical Sciences, Shandong University, Shandong, China.
Anticancer Drugs. 2009 Oct;20(9):800-6. doi: 10.1097/CAD.0b013e32832f9d85.
As the interaction of hyaluronan (HA) with its receptor CD44 contributes to multidrug resistance (MDR) of tumor cells, HA oligosaccharides (o-HAs), as HA antagonists, may be useful to reverse the MDR. The objective of this study was to investigate the reversal effects of four o-HAs, including 4 saccharide residue (o-HA4), 6 saccharide residue (o-HA6), 8 saccharide residue (o-HA8), and 10 saccharide residue (o-HA10) fragments, on adriamycin (ADR)-resistant K562/A02 cells. The four o-HAs were prepared by digesting the native high molecular weight HA with hyaluronidase and gel filtration chromatography. 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay was used to assess the cytotoxicity of the four o-HAs and/or ADR on K562/A02 and K562 cells. The intracellular accumulation of ADR in K562/A02 cells was measured by flow cytometry. By comparing the IC50 (concentration resulting in 50% inhibition of cell growth) of ADR with K562/A02 cells in the presence and absence of a series of different concentrations of o-HAs, the reversal folds of the four o-HAs were calculated. The reversal folds of o-HA4, o-HA6, o-HA8, and o-HA10 were 2.04, 2.05, 1.91, and 1.84, respectively. After o-HA4, o-HA6, o-HA8, and o-HA10 treatment, the intracellular amounts of ADR were increased to 3.90, 3.92, 3.76, and 3.39 times, respectively. Shorter o-HAs (o-HA4 and o-HA6) showed stronger reversal effects than longer o-HAs (o-HA8 and o-HA10). In conclusion, the results showed that the four o-HAs could effectively reverse the ADR resistance of K562/A02 cells by increasing the intracellular accumulation of ADR. O-HAs may be used as MDR reversal drugs to increase the effectiveness of chemotherapy.
由于透明质酸（HA）与其受体CD44的相互作用会导致肿瘤细胞的多药耐药性（MDR），HA寡糖（o-HA）作为HA拮抗剂，可能有助于逆转MDR。本研究的目的是研究四种o-HA，包括4糖残基（o-HA4）、6糖残基（o-HA6）、8糖残基（o-HA8）和10糖残基（o-HA10）片段对阿霉素（ADR）耐药的K562/A02细胞的逆转作用。这四种o-HA是通过用透明质酸酶消化天然高分子量HA并经凝胶过滤色谱法制备的。采用3-[4,5-二甲基噻唑-2-基]-2,5-二苯基溴化四氮唑（MTT）法评估这四种o-HA和/或ADR对K562/A02和K562细胞的细胞毒性。通过流式细胞术测量K562/A02细胞中ADR的细胞内蓄积量。通过比较在存在和不存在一系列不同浓度o-HA的情况下ADR对K562/A02细胞的半数抑制浓度（IC50，即导致细胞生长抑制50%的浓度），计算这四种o-HA的逆转倍数。o-HA4、o-HA6、o-HA8和o-HA10的逆转倍数分别为2.04、2.05、1.91和1.84。经o-HA4、o-HA6、o-HA8和o-HA10处理后，细胞内ADR的量分别增加到3.90、3.92、3.76和3.39倍。较短的o-HA（o-HA4和o-HA6）显示出比长链o-HA（o-HA8和o-HA10）更强的逆转作用。总之，结果表明这四种o-HA可通过增加ADR的细胞内蓄积有效地逆转K562/A02细胞的ADR耐药性。o-HA可用作MDR逆转药物以提高化疗效果。
