不同硫酸软骨素对口腔癌KB细胞的抑制作用

Inhibition Activity of Oral Cancer KB Cell of Chondroitin Sulfate from Different Sources

摘要:

以鲟鱼软骨和鸡软骨为原料，采用碱提-酶解-醇沉的工艺流程提取硫酸软骨素后，利用Sephadex G-50凝胶色谱及Sephacryl S-300 HR凝胶色谱对硫酸软骨素进行纯化，并通过高效凝胶渗透色谱法进行分子质量测定与纯度鉴定。采用MTT法检测硫酸软骨素对口腔癌KB细胞增殖活性的影响，并采用流式细胞术检测口腔癌KB细胞凋亡情况。研究结果表明鲟鱼及鸡软骨硫酸软骨素粗制品得率分别为23.80%和25.23%。经高效液相凝胶渗透色谱分析可得纯化的鲟鱼及鸡软骨硫酸软骨素均为高纯度多糖，且鲟鱼软骨硫酸软骨素的平均分子量比鸡软骨硫酸软骨素的大。以体外抗肿瘤的方法研究硫酸软骨素粗提物和纯化样品对口腔癌KB细胞的抑制作用，结果所得鲟鱼软骨硫酸软骨素粗制品和纯化样品的IC50值分别为80.18 μg/mL和58.78 μg/mL；鸡软骨硫酸软骨素粗制品和纯化样品的IC50值分别为68.55 μg/mL和53.71 μg/mL。流式细胞术表明硫酸软骨素能够诱导KB细胞发生凋亡，细胞凋亡率与药物的浓度呈剂量依赖关系，且鸡软骨硫酸软骨素诱导KB细胞凋亡率高于鲟鱼软骨硫酸软骨素。

Abstract:

Chondroitin sulfate was extracted from the sturgeon cartilage and chicken cartilage using the alkaline extraction-enzymatic-alcohol precipitation process, and purified by Sephadex G-50 gel chromatography and Sephacryl S-300 HR gel chromatography. The molecular mass and purity of the obtained chondroitin sulfate were determined by HPGPC. The effect of chondroitin sulfate on the proliferation of oral cancer KB cells was detected by the MTT assay, and the apoptosis of oral cancer KB cells was examined by flow cytometry. The results showed that the yield of crude chondroitin sulfate from sturgeon cartilage and chicken cartilage was 23.80% and 25.23%, respectively. HPGPC analysis showed that the purified chondrocytes from sturgeon cartilage and chicken cartilage were highly pure polysaccharides, with the average molecular weight of the chondroitin sulfate in sturgeon cartilage higher than that in chicken cartilage. The inhibitory effects of crude and purified chondroitin sulfate on oral cancer KB cells were studied in vitro. The results showed that the IC50 values were 80.18 μg/mL and 58.78 μg/mL, respectively, for the crude and purified chondroitin sulfate from sturgeon cartilage, and 68.55 μg/mL and 53.71 μg/mL, respectively, for the crude and purified chondroitin sulfate from chicken cartilage. Flow cytometry showed that chondroitin sulfate could induce the apoptosis of KB cells, and the apoptosis rate was dependent on the concentration of the chondroitin sulfate. The apoptosis rate induced by the chondroitin sulfate in chicken cartilage was higher than that from sturgeon cartilage.