鼠李柠檬素下调NEK2抑制肝癌细胞恶性生物学行为

Mechanism of rhamnetin inhibiting malignant biological behaviors of hepatocellular carcinoma cells by down-regulating NEK2

目的:分析鼠李柠檬素通过NIMA相关激酶2(NEK2)抑制肝癌细胞增殖、迁移和侵袭的机制。方法:网络药理学分析鼠李柠檬素可能的作用靶点为NEK2,TCGA_LIHC数据库分析肝癌组织中NEK2表达及预后的关系。检测不同浓度鼠李柠檬素对肝癌细胞增殖及NEK2蛋白表达的影响,设置对照组和鼠李柠檬素组,检测两组细胞增殖、迁移和侵袭。观察鼠李柠檬素对移植瘤体积、质量及瘤组织中Ki67蛋白表达。构建shNEK和稳定过表达NEK2细胞,观察鼠李柠檬素抑制NEK2对肝癌细胞生物学行为的影响。结果:与癌旁组织相比,癌组织NEK2的表达明显上调(P<0.05),且其表达量与预后有关(P<0.05)。随着鼠李柠檬素浓度越高,HepG2细胞和Hep3B细胞相对存活率降低(P<0.05)。鼠李柠檬素组增殖活性、侵袭、迁移细胞数均低于对照组(P<0.05),移植瘤体积和质量明显低于对照组(P<0.05),Ki67蛋白表达明显低于对照组(P<0.05),且随着鼠李柠檬素浓度越高、作用时间越久,NEK2蛋白表达下降更为明显(P<0.05)。NEK2基因沉默可以促进细胞增殖、侵袭和迁移,NEK2过表达可以逆转鼠李柠檬素抑制的细胞增殖、侵袭和迁移(P<0.05)。结论:鼠李柠檬素可以抑制NEK2蛋白表达,抑制肝癌细胞的增殖、迁移和侵袭。

Objective:To analyze the mechanism of rhamnetin inhibiting the proliferation,migration and invasion of hepatocellular carcinoma(HCC) cells by NIMA-related kinase 2(NEK2).Methods:The possible target(NEK2) of rhamnetin was analyzed by network pharmacology.According to TCGA_LIHC database,the expression of NEK2 in HCC tissues and its relationship with prognosis were analyzed.The effects of different concentrations of rhamnetin on the proliferation of HCC cells and expression of NEK2 protein were detected.The control group and rhamnetin group were set up.The cells proliferation,migration and invasion were detected.The effect of rhamnetin on the volume and weight of xenograft tumors,and expression of Ki67 protein were observed.The shNEK cells and stable overexpression NEK2 cells were constructed to observe the effects of rhamnetin on biological behaviors of HCC cells by inhibiting NEK2.Results:Compared with para-cancerous tissues,the expression of NEK2 in HCC tissues was up-regulated(P<0.05).With the increase of rhamnetin concentration,relative survival rates of HepG2 and Hep3B were decreased(P<0.05).The proliferation activity,number of invasion and migration cells in rhamnetin group were lower than those in control group(P<0.05),volume and weight of xenograft tumors were lower than those in control group(P<0.05),and the expression of Ki67 protein was significantly lower than that in control group(P<0.05).With the increase of rhamnetin concentration and action time,expression of NEK2 protein was significantly decreased(P<0.05).Silencing NEK2 gene could promote cells proliferation,invasion and migration,while NEK2 overexpression could inhibit cells proliferation,invasion and migration(P<0.05).Conclusion:Rhamnetin can inhibit the expression of NEK2 protein,and inhibit the proliferation,migration and invasion of HCC cells.