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本研究对提取的大黄鱼鱼鳔粗多糖(CPLYCSB)的抗突变效果和对HCT-116结肠癌细胞体外抗癌作用进行了测定。CPLYCSB显示出了对N-甲基-N’-硝基-亚硝基胍(MNNG)和黄曲霉毒素B1(AFB1)诱发鼠伤寒沙门氏菌TA100菌株突变的抗突变效果,浓度为2.5 mg/皿的CPLYCSB比1.25 mg/皿CPLYCSB能够更好的提高抗突变作用。通过MTT(噻唑蓝)法测定出400μg/mL的CPLYCSB处理下HCT-116细胞的生长率降低了72.6%,比200(28.2%)和100μg/mL(12.4%)的CPLYCSB处理时得到了更高的抑制率。相比于低浓度的CPLYCSB,通过DAPI(4,6-二脒基-2-苯基吲哚)染色法观察到400μg/mL的CPLYCSB可以更明显的诱导癌细胞凋亡。400μg/mL高浓度的CPLYCSB可以上调HCT-116癌细胞的caspase-3、-8、-9凋亡基因表达。经过CPLYCSB处理后也可以显著下调炎症相关因子iNOS和COX-2的表达。400μg/mL的CPLYCSB也通过降低HCT-116癌细胞MMP-2和MMP-9基因表达来表现出比其他浓度CPLYCSB更强的抗转移效果。这些体外实验结果证明了CPLYCSB的抗突变和抗癌效果。
In this study, the anti-mutagenic effect of CPLYCSB and the antitumor activity of HCT-116 colon cancer cells in vitro were determined. CPLYCSB showed an anti-mutagenic effect on the mutation of Salmonella typhimurium TA100 strain induced by N-methyl-N’-nitro-nitrosoguanidine (MNNG) and aflatoxin B1 (AFB1) at a concentration of 2.5 mg / CPLYCSB better than 1.25 mg / dish CPLYCSB can improve the anti-mutagenic effect. Growth of HCT-116 cells was reduced by 72.6% at 400 μg / mL CPLYCSB by MTT (thiazolyl blue) assay and higher than CPLYCSB at 200 (28.2%) and 100 μg / mL (12.4%) Inhibition rate. Compared with low concentration of CPLYCSB, 400μg / mL of CPLYCSB was observed to induce cancer cell apoptosis more obviously by DAPI (4,6-diamidino-2-phenylindole) staining. CPLYCSB at a concentration of 400μg / mL could up-regulate the expression of caspase-3, -8, -9 apoptotic genes in HCT-116 cancer cells. After CPLYCSB treatment can significantly down-regulate the expression of inflammation related factors iNOS and COX-2. CPLYCSB at 400 μg / mL also showed stronger anti-metastatic effects than other concentrations of CPLYCSB by reducing MMP-2 and MMP-9 gene expression in HCT-116 cancer cells. These in vitro experimental results demonstrate the anti-mutation and anti-cancer effects of CPLYCSB.