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目的:探讨褐藻糖胶对人肝癌细胞株HepG2体外增殖的相关机制。方法:采用MTT法测定不同浓度褐藻糖胶对人肝癌细胞株HepG2增殖的抑制作用,计算细胞生长抑制率;将0、10、100、500μg/ml的褐藻糖胶作用于HepG2细胞,48 h后光镜观察细胞形态改变,用Hoechst染色法和琼脂糖凝胶电泳检测细胞凋亡;Western blot检测cyclinD1和拓扑异构酶IIα(topoi-somerase IIα,TopoIIα)表达的改变。结果:褐藻糖胶具有抑制人肝癌细胞HepG2增殖的作用,呈剂量依赖性。不同浓度褐藻糖胶作用HepG2细胞后,500μg/ml组较其他浓度组光镜下和Hoechst 33258染色均呈现较明显细胞形态改变,100μg/ml和500μg/ml组均检测出DNA梯形条带。褐藻糖胶也能降低细胞增殖生物标志蛋白cyclinD1和TopoIIα的蛋白表达。结论:褐藻糖胶抑制人肝癌细胞增殖,诱导细胞凋亡,其机制可能与褐藻糖胶抑制细胞增殖的生物标志蛋白cyclinD1和TopoIIα表达有关。
Objective: To investigate the mechanism of fucoidan on the proliferation of human hepatocellular carcinoma cell line HepG2 in vitro. Methods: MTT assay was used to determine the inhibitory effect of different concentrations of fucoidan on the proliferation of HepG2 cells. The cell growth inhibition rate was calculated. Fucoidan at 0, 10, 100 and 500 μg / ml was applied to HepG2 cells for 48 h The morphological changes of cells were observed under light microscope. The apoptosis of cells was detected by Hoechst staining and agarose gel electrophoresis. The expression of cyclinD1 and topoisomerase IIα (TopoIIα) were detected by Western blot. Results: Fucoidan inhibited the proliferation of HepG2 cells in a dose-dependent manner. After HepG2 cells were treated with different concentrations of fucoidan, the morphological changes of cells in 500μg / ml group were more obvious than those in other concentration groups and Hoechst 33258 staining. DNA ladder bands were detected in 100μg / ml and 500μg / ml groups. Fucoidan also decreased the protein expression of the cell proliferation biomarkers cyclinD1 and TopoIIα. CONCLUSION: Fucoidan can inhibit the proliferation of human hepatocellular carcinoma cells and induce the apoptosis of hepatocellular carcinoma cells. The mechanism may be related to the expression of cyclinD1 and TopoIIα, a biomarker of fucoidan that inhibits cell proliferation.