鲟鱼硫酸软骨素对结直肠癌细胞抑制作用

Inhibitory Effect of Chondroitin Sulfate from Sturgeon Bone on Colorectal Cancer Cells

探究鲟鱼硫酸软骨素对结直肠癌细胞的生长抑制能力,并初步确定影响结直肠癌细胞生长的主要原因。选取3株具有代表性的结直肠癌细胞Caco-2、HCT-116、SW480,采用CCK-8法测定细胞生长曲线、硫酸软骨素对癌细胞增殖的抑制能力;以其中一株结直肠癌细胞HCT-116为模型,用流式细胞仪测定其周期和凋亡情况,用细胞核染色法观察处理前后细胞形态,结合caspase-3凋亡酶活力的测定,考察鲟鱼硫酸软骨素对结直肠癌细胞的促凋亡能力,并初步确定作用途径。研究结果表明,鲟鱼硫酸软骨素对结直肠癌细胞Caco-2、HCT-116、SW480的增殖具有一定抑制作用,最高抑制率分别为70.94%、90.00%和75.00%,明显高于阳性对照处理组;处理后,细胞周期G1/G0期比例升高至88.56%,S期比例降低至4.47%,阻滞细胞G1/G0期;同时,使用鲟鱼硫酸软骨素处理细胞后,细胞形态发生变化,出现细胞核固缩以及细胞破碎等现象,细胞主要的凋亡酶活力显著升高,酶活力可达1 645 IU/μg,使细胞发生凋亡,凋亡率可达63.73%。研究结果证明,鲟鱼硫酸软骨素具有促进结直肠癌细胞凋亡、抑制细胞增殖的能力。

Objective: This study is aimed to investigate the inhibitory effect of chondroitin sulfate from sturgeon bone on the proliferation of colorectal cancer cells and to elucidate the underlying mechanism.Methods: Three representative colorectal cancer cells(Caco-2,HCT-116,and SW480) were used in this study.Cell growth and proliferation was tested by cell counting kit-8(CCK-8) method.Colorectal cancer cell line HCT-116 was used to further investigate the pro-apoptotic ability of sturgeon chondroitin sulfate.Cell cycle and apoptosis were determined by flow cytometry.Cell morphology was observed after cell staining.Apoptosis was evaluated by caspase-3 activity.Our study indicated that sturgeon chondroitin sulfate significantly inhibit the cell proliferation of Caco-2,HCT-116 and SW480 cells with a percentage suppression of 70.94%,90.00% and 75.00%,respectively.After sturgeon chondroitin sulfate treatment,the G1/G0 phase increased to 88.56% and the S phase declined to 4.47%.Meanwhile,sturgeon chondroitin sulfate-treated cells exhibited morphological changes,nuclear pyknosis and cell fragmentation,and significantly increased apoptotic enzyme activity(1 645 IU/μg) with an apoptosis rate of 63.73%.Sturgeon chondroitin sulfate showed potential anticancer activity,which will make it a new functional food ingredient.