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目的探讨花蕊石对人肝癌SMMC-7721细胞增殖抑制作用及其初步作用机制。方法采用CCK-8法检测花蕊石含药血清对肝癌细胞SMMC-7721的增殖抑制作用;Annexin V-FITC/PI双染法检测细胞凋亡;Western blotting法检测PTEN/PI3K/AKT信号通路中第10号染色体缺失的磷酸酶及张力蛋白同源的基因(PTEN)、磷脂酰肌醇3-激酶(PI3K)和磷酸蛋白激酶B(p-Akt)相关蛋白的表达水平;RT-PCR检测肿瘤坏死因子α(TNF-α)及核因子活化B细胞κ轻链增强子(NF-κB)相关基因mRNA的表达水平。结果花蕊石具有明显的抑制肝癌SMMC-7721细胞增殖及诱导细胞凋亡的作用。花蕊石含药血清可明显上调PTEN/PI3K/AKT信号通路中PTEN蛋白的表达水平,降低PI3K及p-Akt蛋白的表达,同时下调NF-κB mRNA的表达。结论花蕊石能够抑制肝癌SMMC-7721细胞增殖,并促进肝癌细胞凋亡,其作用机制可能与激活PTEN表达水平从而抑制PI3K/AKT信号通路及其下游基因NF-κB的异常活化有关。
Objective To investigate the inhibitory effect of flower bud stone on the proliferation of human hepatoma SMMC-7721 cells and its primary mechanism. Methods The proliferation of SMMC-7721 cells was detected by CCK-8 assay. The apoptosis of SMMC-7721 cells was detected by Annexin V-FITC / PI double staining. The expression of PTEN / PI3K / AKT was detected by Western blotting (PTEN), phosphatidylinositol 3-kinase (PI3K) and phosphorylated protein kinase B (p-Akt) related proteins in chromosome 10 were detected by RT-PCR. Tumor necrosis (TNF-α) and nuclear factor kappa B (NF-κB) -related gene mRNA expression levels in activated B cells. Results Flower stone has a significant inhibitory effect on proliferation and induction of apoptosis in hepatoma SMMC-7721 cells. The antisense serum-containing serum could significantly up-regulate the expression of PTEN protein in PTEN / PI3K / AKT signaling pathway, decrease the expression of PI3K and p-Akt protein, and down-regulate the expression of NF-κB mRNA. Conclusion Flower bud stone can inhibit the proliferation of hepatocellular carcinoma SMMC-7721 cells and promote the apoptosis of hepatocellular carcinoma cells. The mechanism may be related to the activation of PTEN expression and the inhibition of PI3K / AKT signaling pathway and the abnormal activation of its downstream gene NF-κB.