基于PI3K/mTOR通路益气化瘀解毒方对人肝癌细胞SMCC-7721自噬流的影响(1)
摘要：目的 观察益气化瘀解毒方对人肝癌细胞SMCC-7721自噬流的影响，初步探讨其可能的作用机制。方法 实验分为空白对照组、重楼皂苷Ⅰ组、益气化瘀解毒方组、雷帕霉素组及索拉非尼组，各给药组给予相应药物。体外实验观察人肝癌细胞SMCC-7721自噬流发生情况并检测细胞微管相关蛋白1轻链3(LC-3Ⅱ)、类胰岛素样生长因子2的mRNA结合蛋白质(p62)及磷脂酰肌醇-3-激酶(PI3K)、哺乳动物雷帕霉素靶蛋白(mTOR)的表达。结果 重楼皂苷Ⅰ、益气化瘀解毒方及索拉非尼均能阻断人肝癌细胞SMCC-7721自噬流，导致细胞内自噬小体的蓄积，上调LC-3Ⅱ、p62的表达，同时可下调PI3K、mTOR蛋白的表达。结论 益气化瘀解毒方可改变人肝癌细胞SMCC-7721形态;重楼皂苷Ⅰ、益气化瘀解毒方及索拉非尼均能阻断人肝癌细胞SMCC-7721自噬流，增加自噬体数量，其机制可能与抑制PI3K/mTOR信号通路有关。
关键词：益气化瘀解毒方;自噬流;人肝癌细胞SMCC-7721
中图分类号：R285.5 文献标识码：A 文章编号：1005-5304(2019)10-0045-05
DOI：10.3969/j.issn.1005-5304.2019.10.011 开放科学(资源服务)标识码(OSID)：
Abstract： Objective To observe the effects of Yiqi Huayu Jiedu Prescription on autophagy flow of human hepatocellular carcinoma cell SMCC-7721; To preliminarily discuss its possible mechanism of action. Methods The experiment was divided into blank control group， Chonglou saponin Ⅰ group， Yiqi Huayu Jiedu Prescription group， Rapamycin group and Sorafenib group. Each administration group was given relevant medicine. In vitro experiments were performed to observe the occurrence of autophagy flow in human hepatocellular carcinoma cell SMCC-7721 and to detect the expressions of microtubule-associated protein 1 light chain 3 (LC-3Ⅱ)， insulin-like growth factor 2 mRNA-binding protein (p62) and phosphatidylinositol-3-kinase (PI3K)， mammalian target of Rapamycin (mTOR). Results Chonglou Saponins Ⅰ， Yiqi Huayu Jiedu Prescription and Sorafenib could block the autophagy flow of human hepatocellular carcinoma cell SMCC-7721， leading to the accumulation of autophagic bodies in cells and up-regulating the expression of LC-3Ⅱ and p62. The expressions of PI3K and mTOR protein could be down-regulated. Conclusion Yiqi Huayu Jiedu Prescription can change cell morphology of SMCC-7721; Chonglou saponins Ⅰ， Yiqi Huayu Jiedu Prescription， and sorafenib can block the autophagy flow of SMCC-7721 and increase the number of autophagic bodies， which may be related to the PI3K/mTOR signal pathway.
Keywords： Yiqi Huayu Jiedu Prescription; autophagy flow; human hepatocellular carcinoma cell SMCC-7721
據统计，原发性肝癌发病率在恶性肿瘤中居第6位，死亡率居第4位，我国肝癌患者死亡人数占全球一半以上[1]。中医药资源丰富，是治疗原发性肝癌的手段之一，长期服用毒副作用低[2]。有研究表明，中药在抑制血管新生、抗凋亡、调节免疫及改善生活质量等方面发挥了重要的作用[3-4]。益气化瘀解毒方是全国名中医潘敏求教授治疗肝癌效验方。我们前期研究发现，其可下调人肝癌细胞CXCL12/CXCR4/ CXCR7趋化因子轴，抑制细胞增殖及迁移，同时能降低肝癌裸鼠移植瘤血管密度(MVD)，抑制低氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)的表达，从而发挥抗肿瘤作用[5-10]。下调HIF-1α能通过多种途径促进VEGF的表达，VEGF是PI3K/mTOR信号通路上游的关键靶点之一，且PI3K/mTOR信号通路是参与细胞自噬的经典途径之一。因此，在前期实验的基础上，本研究以PI3K/mTOR信号通路介导肝癌细胞自噬性死亡为可能机制，进一步探讨益气化瘀解毒方抗肿瘤的作用途径，为开发抗肝癌多靶点的中药新药提供依据。, 百拇医药(张振 郜文辉 田雪飞 陈思勤 何凤姣 周芳 王亚琪 朱洪兵 李克雄 曾普华)
关键词：益气化瘀解毒方;自噬流;人肝癌细胞SMCC-7721
中图分类号：R285.5 文献标识码：A 文章编号：1005-5304(2019)10-0045-05
DOI：10.3969/j.issn.1005-5304.2019.10.011 开放科学(资源服务)标识码(OSID)：
Abstract： Objective To observe the effects of Yiqi Huayu Jiedu Prescription on autophagy flow of human hepatocellular carcinoma cell SMCC-7721; To preliminarily discuss its possible mechanism of action. Methods The experiment was divided into blank control group， Chonglou saponin Ⅰ group， Yiqi Huayu Jiedu Prescription group， Rapamycin group and Sorafenib group. Each administration group was given relevant medicine. In vitro experiments were performed to observe the occurrence of autophagy flow in human hepatocellular carcinoma cell SMCC-7721 and to detect the expressions of microtubule-associated protein 1 light chain 3 (LC-3Ⅱ)， insulin-like growth factor 2 mRNA-binding protein (p62) and phosphatidylinositol-3-kinase (PI3K)， mammalian target of Rapamycin (mTOR). Results Chonglou Saponins Ⅰ， Yiqi Huayu Jiedu Prescription and Sorafenib could block the autophagy flow of human hepatocellular carcinoma cell SMCC-7721， leading to the accumulation of autophagic bodies in cells and up-regulating the expression of LC-3Ⅱ and p62. The expressions of PI3K and mTOR protein could be down-regulated. Conclusion Yiqi Huayu Jiedu Prescription can change cell morphology of SMCC-7721; Chonglou saponins Ⅰ， Yiqi Huayu Jiedu Prescription， and sorafenib can block the autophagy flow of SMCC-7721 and increase the number of autophagic bodies， which may be related to the PI3K/mTOR signal pathway.
Keywords： Yiqi Huayu Jiedu Prescription; autophagy flow; human hepatocellular carcinoma cell SMCC-7721
據统计，原发性肝癌发病率在恶性肿瘤中居第6位，死亡率居第4位，我国肝癌患者死亡人数占全球一半以上[1]。中医药资源丰富，是治疗原发性肝癌的手段之一，长期服用毒副作用低[2]。有研究表明，中药在抑制血管新生、抗凋亡、调节免疫及改善生活质量等方面发挥了重要的作用[3-4]。益气化瘀解毒方是全国名中医潘敏求教授治疗肝癌效验方。我们前期研究发现，其可下调人肝癌细胞CXCL12/CXCR4/ CXCR7趋化因子轴，抑制细胞增殖及迁移，同时能降低肝癌裸鼠移植瘤血管密度(MVD)，抑制低氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)的表达，从而发挥抗肿瘤作用[5-10]。下调HIF-1α能通过多种途径促进VEGF的表达，VEGF是PI3K/mTOR信号通路上游的关键靶点之一，且PI3K/mTOR信号通路是参与细胞自噬的经典途径之一。因此，在前期实验的基础上，本研究以PI3K/mTOR信号通路介导肝癌细胞自噬性死亡为可能机制，进一步探讨益气化瘀解毒方抗肿瘤的作用途径，为开发抗肝癌多靶点的中药新药提供依据。, 百拇医药(张振 郜文辉 田雪飞 陈思勤 何凤姣 周芳 王亚琪 朱洪兵 李克雄 曾普华)