健脾益气方含药血清对人肝癌细胞SMMC-7721增殖与侵袭能力的影响

Effects of medicated serum containing Jianpi Yiqi decoction on proliferation and invasion of hepatic carcinoma cells SMMC-7721

目的:观察健脾益气方含药血清对人肝癌细胞SMMC-7721增殖与侵袭能力的影响。方法:以TGF-β1诱导人肝癌细胞SMMC-7721上皮-间质转化(EMT)模型,制备健脾益气方大鼠含药血清,分别以0%、5%、10%、15%、20%、30%含药血清浓度干预人肝癌细胞SMMC-7721 12 h、24 h、48 h、72 h后,采用四甲基偶氮唑盐(MTT)比色法检测细胞的增殖能力,采用Transwell细胞体外侵袭实验观察细胞的侵袭能力。结果:健脾益气方对SMMC-7721 EMT细胞的增殖有抑制作用,且随含药血清浓度增高而递增,呈时间依赖性;12 h 10%含药血清,24 h 15%、20%、30%含药血清,48 h 15%含药血清与相对应浓度空白血清比较,均具有显著性差异(P<0.01),其中以48 h 15%含药血清浓度对细胞增殖的抑制效果最明显。健脾益气方含药血清各个浓度组与对照组相比,细胞侵袭能力均有不同程度的下降,其中10%、15%、20%、30%浓度的含药血清都具有显著性差异(P<0.01);而与空白血清组相比,15%、20%、30%浓度的含药血清均具有显著性差异(P<0.01),其中以15%浓度的含药血清对细胞侵袭的抑制作用最明显。结论:15%健脾益气方含药血清作用48 h,对人肝癌细胞SMMC-7721EMT模型增殖与侵袭的抑制效果最好。

Objective： To observe effects of medicated serum containing Jianpi Yiqi decoction on proliferation and invasion of hepatic carcinoma cells SMMC-7721. Methods： The epithelial- mesenchymal transition（EMT）model of hepatic carcinoma cells was induced by TGF-β_1 to make medicated serum containing Jianpi Yiqi decoction. After 12 h, 24 h, 48 h, 72 h, TGF-β1-induced SMMC-7721 cells were interfered with medicated serum with the concentration of 0%,5%,10%,15%,20%,30%,respectively. The proliferation effect on SMMC-7721 was measured by MTT colorimetric assay and the invasion ability was observed by Transwell invasion assay in vitro.Results： Jianpi Yiqi decoction had inhibitory effect on proliferation for SMMC-7721 EMT cell, which strengthened with the increase of concentration of medicated serum and exhibited time-dependence. Compared with the corresponding blank serum, the inhibition effect of medicated serum with the concentration of 10% on cell proliferation had significant difference at 12 hours（P〈0.01）, as well as the medicated serum with the concentration of 15%,20%,30% at 24 hours, and 15% at 48 hours. The medicated serum with the concentration of 15% had the most obvious inhibition effect on cell proliferation at 48 hours. The invasive ability of SMMC-7721 cells was significantly decreased to varying degree after treatment with different concentrations of medicated serum compared with that of control group. The medicated serum with the concentration of 10%,15%,20%,and 30% had significant difference（P〈0.01）. Compared with the corresponding blank serum, medicated serum with the concentration of 15%,20%, 30% had statistic significance（P 0.01）. The medicated serum with the concentration of 15% had the most obvious inhibition effect on cell invasion. Conclusion： The medicated serum with the concentration of 15% at 48 hours has the best inhibition effect on proliferation and invasion of SMMC-7721 EMT model of hepatic carcinoma cells.