安罗替尼通过PI3K/Akt信号通路调控人肝癌细胞株HepG2增殖迁移的机制OA
Mechanism of Anlotinib in regulating the proliferation and migration of human hepatocellular carcinoma cell line HepG2 through the PI3K/Akt signaling pathway
目的 探究安罗替尼对人肝癌细胞增殖与迁移产生的影响,同时判断其是否借助PI3K/Akt信号通路来发挥作用.方法 安罗替尼以及PI3K抑制剂LY294002应用到经过传代培养的人肝癌细胞株HepG2上.实验设置了安罗替尼与PI3K抑制剂LY294002联合用药的组别,同时设有对照组.为检测细胞的增殖、克隆形成以及迁移能力,分别采用了 CCK-8法和细胞划痕愈合实验.此外,运用蛋白质印迹法来检测安罗替尼和LY294002对人肝癌HepG2细胞中基质金属蛋白酶-3(MMP-3)以及磷酸化Akt(p-Akt)蛋白表达产生的影响.结果 人肝癌细胞株HepG2经安罗替尼作用24 h后,最合适的药物用量是5 000 nmol/L,此时细胞抑制率达到了 49.649 9％.与对照组相比,5 000 nmol/L安罗替尼处理组和8 μmol/L LY294002处理组的人肝癌HepG2细胞增殖率均较低,差异有统计学意义(P＜0.05).在克隆形成和迁移实验中,安罗替尼组、LY294002组以及两药联合组的克隆形成率和迁移率均低于对照组,差异有统计学意义(P＜0.05);并且,两药联合组的克隆形成率和迁移率均分别低于安罗替尼单独处理组和LY294002单独处理组,差异有统计学意义(P＜0.05).在蛋白表达方面,安罗替尼组、LY294002组及联合组的p-Akt和MMP-3蛋白表达水平均低于对照组,差异有统计学意义(P＜0.05);同时,联合用药组中p-Akt蛋白和MMP-3蛋白的表达水平均分别低于安罗替尼单独处理组和LY294002单独处理组,差异有统计学意义(P＜0.05).结论 安罗替尼对肝癌HepG2细胞的增殖与迁移具备抑制作用,这种抑制效果的作用机理或许和对PI3K/Akt信号通路的抑制存在关联.
Objective To investigate the impact of Anlotinib on the proliferation and migration of human hepatocellular car-cinoma cells and to determine whether its mechanism involves the PI3K/Akt signaling pathway.Methods Anlotinib and the PI3K inhibitor LY294002 were applied to passaged human hepatocellular carcinoma cell line HepG2.Experimental groups included Anlotinib alone,the PI3K inhibitor LY294002 alone,a combination of both drugs,and a control group.Cell proliferation,clonogenic formation,and migration capabilities were assessed using the CCK-8 assay and the cell scratch wound healing assay,respectively.Additionally,Western blotting was employed to examine the effects of Anlotinib and LY294002 on the expression of matrix metalloproteinase-3(MMP-3)and phosphorylated Akt(p-Akt)proteins in HepG2 cells.Results After 24 hours of treatment with Anlotinib,the optimal concentration for HepG2 cells was determined to be 5 000 nmol/L,achieving a cell inhibition rate of 49.649％.Compared with the control group,the proliferation rates of HepG2 cells in both the 5 000 nmol/L Anlotinib treatment group and the 8 μmol/L LY294002 treatment group were significantly lower,the differences were statistically significant(P＜0.05).In the clonogenic formation and migration as-says,the clonogenic formation rates and migration rates of the Anlotinib group,the LY294002 group,and the combination group were all lower than those of the control group,with statistically significant differences(P＜0.05).Furthermore,the clonogenic formation rates and migration rates in the combination group were lower than those in either the Anlotinib alone group or the LY294002 alone group,the differences were statistically significant(P＜0.05).Regarding protein expression,the expression levels of p-Akt and MMP-3 proteins in the anlotinib group,LY294002 group,and combination group were lower than those in the control group,the differences were statistically significant(P＜0.05).Additionally,the expression levels of both p-Akt and MMP-3 proteins in the combination group were significantly lower than those in either the Anlotinib alone group or the LY294002 alone group,the differences were statistically significant(P＜0.05).Conclusion Anlotinib exhibits inhibitory effects on the proliferation and migration of hepatocellular carcinoma HepG2 cells.The underlying mechanism of this inhibitory effect may be associated with the suppression of the PI3K/Akt signaling pathway.
张静宇;张鸿雁;赵晶;杨洁;齐云飞;孟祥海
菏泽医学专科学校基础医学部组织学与胚胎学教研室,山东菏泽 274000菏泽医学专科学校基础医学部组织学与胚胎学教研室,山东菏泽 274000菏泽医学专科学校教务科,山东菏泽 274000菏泽医学专科学校基础医学部组织学与胚胎学教研室,山东菏泽 274000菏泽医学专科学校基础医学部组织学与胚胎学教研室,山东菏泽 274000山东省鄄城县中医医院康复科,山东菏泽 274600
医药卫生
安罗替尼LY294002蛋白质金属基质-3细胞增殖HepG2
AnlotinibLY294002Matrix metalloproteinase-3Cell proliferationHepG2
《中国当代医药》 2026 (11)
4-10,7
山东省医药卫生科技发展计划项目(202103101086).
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