南方医科大学学报

南方医科大学学报 ›› 2019, Vol. 39 ›› Issue (10): 1180-.doi: 10.12122/j.issn.1673-4254.2019.10.08

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白术多糖对肝癌细胞增殖及侵袭的抑制作用及其机制

朱云，李成，林鑫盛，孙晶晶，程旸   

  • 出版日期:2019-10-20 发布日期:2019-10-20

Effect of Atractylodes macrocephala polysaccharide on proliferation and invasion of hepatocellular carcinoma cells in vitro

  • Online:2019-10-20 Published:2019-10-20

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摘要： 目的探讨白术多糖（PAM）对肝癌细胞增殖和侵袭能力的影响及其机制。方法体外培养肝癌细胞HepG2分别给予不同 浓度的PAM处理，通过CCK-8和Transwell实验检测肝癌细胞的增殖、侵袭能力。采用免疫荧光技术检测肝癌细胞中β-catenin 蛋白表达水平，采用RT-PCR技术和免疫印迹法检测AKT、GSK-3β的基因和蛋白表达水平及其磷酸化表达，检测MMP-2的基 因和蛋白表达水平。HepG2 细胞给予LiCl/PAM/LiCl 联合PAM处理，检测细胞增殖、侵袭能力改变，检测GSK-3β磷酸化和 MMP2的蛋白水平。结果与空白对照组相比，PAM处理后肝癌细胞体外增殖能力下降、侵袭能力下降（P<0.05）；PAM处理可 以下调β-catenin蛋白表达、下调MMP-2基因与蛋白表达水平，同时抑制AKT与GSK-3β的磷酸化（P<0.05）。PAM对肝癌细胞 体外增殖、侵袭及AKT/GSK-3β/β-catenin通路的影响呈浓度依赖性：PAM的浓度越高，对肝癌细胞体外增殖、侵袭的抑制作用 越强；对AKT与GSK-3β的磷酸化的抑制作用越强，对β-catenin表达的抑制作用也越强。LiCl可以逆转PAM对肝癌细胞增殖、 侵袭的抑制作用，同时可以逆转PAM对GSK-3β磷酸化和MMP2的蛋白表达的抑制。结论PAM对肝癌细胞的体外增殖和侵 袭能力具有抑制作用，PAM可能通过影响Wnt/β-catenin 信号通路发挥作用。

Abstract: Objective To investigate the inhibitory effect of polysaccharide of Atractylodes macrocephala (PAM) on the proliferation and invasion of hepatocellular carcinoma cells and the underlying mechanism. Methods Hepatocellular carcinoma HepG2 cells were treated with different concentrations of PAM, and their proliferation and invasive ability were examined using CCK-8 assay and Transwell assay. Immunofluorescence assay was performed to detect the expression level of β-catenin, and real-time PCR and Western blotting were used to detect the mRNA and protein expressions of AKT, GSK-3β and MMP-2 in the cells. The changes in the proliferation, invasiveness and the expressions of pGSK-3β and MMP2 were examined in the cells following treatment with LiCl/PAM/LiCl plus PAM. Results PAM treatment significantly reduced the cell viability, the number of migration cells, and the expression levels of β-catenin and MMP-2 (P<0.05), and obviously inhibited the phosphorylation of AKT and GSK-3β in the cells (P<0.05) in a dose-dependent manner. The rescue experiment showed that LiCl reversed the inhibition of cell proliferation, invasiveness, and the Wnt/β-catenin pathway induced by PAM. Conclusion PAM can inhibit the proliferation and invasion of hepatocellular carcinoma cells in vitro possibly by inhibiting the Wnt/β-catenin signaling pathway.