健脾1号方对人结肠癌细胞作用及其机制研究

Effect and Mechanism of No.1 Jianpi Prescription on Human Colon Cancer HCT116 Cells

目的:探讨健脾1号方抑制人结肠癌细胞增殖和干细胞特性的作用机制。方法:以人结肠HCT116癌细胞株为模型,通过MTT实验检测不同浓度(0.4、2、10及50 mg/m L)健脾1号方对HCT116细胞的增殖抑制作用;以无血清悬浮培养实验检测健脾1号方对HCT116细胞干细胞特性的影响;通过Western-blot检测健脾1号方对HCT116细胞钙黏附蛋白E(E-cadherin)表达的影响。结果:健脾1号方对HCT116生长具有抑制作用,且导致细胞形态发生变化,干预24、48及72h后的IC50值分别为2.32、1.78、1.31 mg/m L;健脾1号方具有抑制HCT116细胞成球能力,上调E-cadherin蛋白表达。结论:健脾1号方对HCT116细胞生长和干细胞特性具有明显的抑制作用,其机制可能和调节细胞上皮间质转化(EMT)有关。

Objective： To investigate the effect of No.1 Jianpi Prescription on growth inhibition and cancer stem celllike property in HCT116 cells and to explore the related molecular mechanism. Methods： Human colon cancer HCT116 cells cultured in vitro were treated with different concentrations of No. 1 Jianpi Prescription（ 0. 4 mg / m L,2 mg / m L,10 mg / m L,50 mg / m L） for 24 h,48 h and 72 h. MTT assay was used to determine cells proliferation of HCT116 cells. The self-renewal capacity was determined by tumorshphere formation assay. The expressions of E-cadherin were examined by Western blot. Results： The cell growth was significantly inhibited by No.1 Jianpi Prescription in HCT116 cells with the IC50 values were 2.32 mg / m L,1.78 mg / m L,1.31 mg / m L in 24 h,48 h,72 h,respectively. Cell morphology changes were observed by optical microscope. The self-renewal capacity of HCT116 cells was inhibited by No.1 Jianpi Prescription. Furthermore,No.1 Jianpi Prescription increased the expression of E-cadherin in HCT116 cells. Conclusion： No. 1Jianpi Prescription can inhibit the cell proliferation and self-renewal of HCT116 cells. This inhibitory effect may be related with suppression of EMT.