健脾解毒方对过表达HBx肝癌HepG2细胞PI3K/AKT通路的影响(1)
摘要：目的 观察健脾解毒方对过表达乙型肝炎病毒X蛋白(HBx)的肝癌细胞增殖及PI3K/AKT信号通路相关因子表达的影响。方法 构建过表达HBx肝癌的HepG2细胞，给予不同浓度健脾解毒方药物血清进行干预，实验分为肝癌细胞空白组、pcDNA3.1组、pcDNA3.1-HBx组和健脾解毒方低、中、高剂量组。采用MTT法及流式细胞仪检测细胞增殖及凋亡水平，采用Western blot检测肝癌细胞p-PI3K、p-AKT蛋白表达。结果 成功构建过表达HBx的肝癌HepG2细胞，发现有促进肝癌细胞增殖及上调p-PI3K及p-AKT表达的作用。健脾解毒方药物血清干预后，可不同程度抑制肝癌细胞增殖、促进肝癌细胞凋亡，以及下调p-AKT、p-PI3K表达的作用，而健脾解毒方低剂量组对p-PI3K表达作用不显著。结论 健脾解毒方有抑制肝癌细胞增殖及促进凋亡的作用，其机制与下调PI3K/AKT通路相关因子表达有关。
关键词：肝癌细胞；PI3K/AKT信号通路；乙型肝炎病毒X蛋白；健脾解毒方
中图分类号：R285.5 文献标识码：A 文章编号：1005-5304(2019)03-0056-04
Abstract： Objective To study the effects of Jianpi Jiedu Prescription on proliferation of HBx overexpression hepatoma cells and expression of PI3K/AKT signaling pathway-related factors. Methods HepG2 cells of HBx overexpression were established. Different concentrations of Jianpi Jiedu Prescription serum were given for intervention. The study was divided into hepatoma cells blank group， pcDNA3.1 group， pcDNA3.1-HBx group and Jianpi Jiedu Prescription low-， medium- and high-dosage groups. Cell proliferation and apoptosis were detected by MTT assay and flow cytometry. The expressions of p-PI3K and p-AKT protein were detected by Western blot. Results HepG2 cells of HBx overexpression were successfully constructed. It was found that HBx overexpression could promote the proliferation of hepatoma cells and up-regulate the expressions of p-PI3K and p-AKT. After intervention with serum of Jianpi Jiedu Prescription， it was found that each Jianpi Jiedu Prescription group inhibited the proliferation of hepatoma cells， promoted apoptosis of hepatoma cells and decreased the expressions of p-AKT and p-PI3K， but Jianpi Jiedu Prescription low-dosage group had no significant effect on p-PI3K. Conclusion Jianpi Jiedu Prescription has the effect of inhibiting the proliferation and promoting apoptosis of hepatoma cells， and its mechanism is related to down-regulating the expression of PI3K/AKT pathway-related factors.
Keywords： hepatoma cells； PI3K/AKT signaling pathway； HBx； Jianpi Jiedu Prescription
原發性肝癌(primary liver cancer，PLC)是发病率较高的恶性肿瘤，其全球发病率和死亡率分别居所有肿瘤的第7位和第4位[1]，治疗难度极大，预后很差。目前PLC机制尚不清楚，因而加强其发病机制研究，并寻找有效治疗手段是肝癌防治的关键所在[2-3]。PI3K/AKT信号通路在肝癌细胞生长、分化、增殖、迁移和存活中扮演重要角色，与肝癌发生发展密切相关[4-5]。乙型肝炎病毒(hepatitis B virus，HBV)慢性感染是肝细胞癌发病的独立高危因素，其中乙型肝炎病毒X蛋白(HBx)是一种重要的反式作用因子，能广泛激活多种原癌基因的转录表达，是肝细胞恶性转化的重要原因，被认为在肝癌发生过程中发挥直接促癌作用[6-8]。我们前期通过二甲基亚硝胺诱导大鼠肝癌模型，采用健脾解毒方对其干预后，发现有延缓肝癌发病的作用，其机制与中药下调PI3K/AKT信号通路相关因子表达有关[9-10]。为进一步探索其机制，本研究构建过表达HBx的肝癌HepG2细胞，采用健脾解毒方药物血清进行干预，观察其对过表达HBx的肝癌细胞增殖及PI3K/AKT信号通路相关因子表达的影响。, 百拇医药(张斌 王晓薇 徐春江 丁慎华 朱薇珊 钱雪梅)
关键词：肝癌细胞；PI3K/AKT信号通路；乙型肝炎病毒X蛋白；健脾解毒方
中图分类号：R285.5 文献标识码：A 文章编号：1005-5304(2019)03-0056-04
Abstract： Objective To study the effects of Jianpi Jiedu Prescription on proliferation of HBx overexpression hepatoma cells and expression of PI3K/AKT signaling pathway-related factors. Methods HepG2 cells of HBx overexpression were established. Different concentrations of Jianpi Jiedu Prescription serum were given for intervention. The study was divided into hepatoma cells blank group， pcDNA3.1 group， pcDNA3.1-HBx group and Jianpi Jiedu Prescription low-， medium- and high-dosage groups. Cell proliferation and apoptosis were detected by MTT assay and flow cytometry. The expressions of p-PI3K and p-AKT protein were detected by Western blot. Results HepG2 cells of HBx overexpression were successfully constructed. It was found that HBx overexpression could promote the proliferation of hepatoma cells and up-regulate the expressions of p-PI3K and p-AKT. After intervention with serum of Jianpi Jiedu Prescription， it was found that each Jianpi Jiedu Prescription group inhibited the proliferation of hepatoma cells， promoted apoptosis of hepatoma cells and decreased the expressions of p-AKT and p-PI3K， but Jianpi Jiedu Prescription low-dosage group had no significant effect on p-PI3K. Conclusion Jianpi Jiedu Prescription has the effect of inhibiting the proliferation and promoting apoptosis of hepatoma cells， and its mechanism is related to down-regulating the expression of PI3K/AKT pathway-related factors.
Keywords： hepatoma cells； PI3K/AKT signaling pathway； HBx； Jianpi Jiedu Prescription
原發性肝癌(primary liver cancer，PLC)是发病率较高的恶性肿瘤，其全球发病率和死亡率分别居所有肿瘤的第7位和第4位[1]，治疗难度极大，预后很差。目前PLC机制尚不清楚，因而加强其发病机制研究，并寻找有效治疗手段是肝癌防治的关键所在[2-3]。PI3K/AKT信号通路在肝癌细胞生长、分化、增殖、迁移和存活中扮演重要角色，与肝癌发生发展密切相关[4-5]。乙型肝炎病毒(hepatitis B virus，HBV)慢性感染是肝细胞癌发病的独立高危因素，其中乙型肝炎病毒X蛋白(HBx)是一种重要的反式作用因子，能广泛激活多种原癌基因的转录表达，是肝细胞恶性转化的重要原因，被认为在肝癌发生过程中发挥直接促癌作用[6-8]。我们前期通过二甲基亚硝胺诱导大鼠肝癌模型，采用健脾解毒方对其干预后，发现有延缓肝癌发病的作用，其机制与中药下调PI3K/AKT信号通路相关因子表达有关[9-10]。为进一步探索其机制，本研究构建过表达HBx的肝癌HepG2细胞，采用健脾解毒方药物血清进行干预，观察其对过表达HBx的肝癌细胞增殖及PI3K/AKT信号通路相关因子表达的影响。, 百拇医药(张斌 王晓薇 徐春江 丁慎华 朱薇珊 钱雪梅)