莼菜粗多糖对Hela细胞生长影响的初步研究

Preliminary Study on the Effect of Crude Polysaccharide from Brasenia schreberi on HeLa Cell Growth

摘要: 为探究莼菜粗多糖对Hela细胞生长的抑制作用，本研究以湖北省利川市莼菜嫩芽为原材料，提取粗多糖(BSP)，通过MTT法测抑制率，AO-PI双染观察细胞，对不同非正常细胞形态进行计数，用RT-qPCR方法检测部分癌症相关基因表达情况。在2000个/孔细胞初始密度时，500 µg/mL BSP就对Hela细胞有抑制作用，2000 µg/mL BSP处理Hela细胞72 h，抑制率可达到96.88%。BSP诱导Hela细胞产生四种非正常形态，随BSP浓度升高，I型比例减小，II、III、IV型比例增大。250 µg/mL BSP处理Hela细胞12 h，p53、Cyclin D1、CDK2和CDK7基因相对表达水平均在2倍左右。BSP对Hela细胞的抑制作用呈时间浓度依赖，且会破坏细胞膜，改变细胞形态。抑制作用可能与p53及细胞周期等调控基因的积累有关。

Abstract: To explore the inhibitory effect of crude Brasenia schreberi polysaccharides on Hela cells, the crude polysaccharide (BSP) was extracted from the shoots of Brasenia schreberi in Lichuan City, Hubei Province. The inhibition rate was measured by MTT method, the cells were observed by AO-PI double staining, the different abnormal cell morphology was counted, and the expression of some cancer-related genes was detected by RT-qPCR. At the initial density of 2000 cells/well, Hela cells were inhibited by 500 μg/mL BSP. With the increase of BSP concentration, the proportion of type I decreased and the proportion of type II, III and IV increased. When HeLa cells were treated with 250 μg/mL BSP for 12 hours, the relative expression levels of p53, Cyclin D1, CDK2 and CDK7 genes were about 2-fold of the control. The inhibitory effect of BSP on HeLa cells was time-dependent and concentration-dependent, and it would destroy cell membrane and change cell morphology. The inhibition may be related to the accumulation of p53 and cyclin.