槐耳上清对人胃癌HGC-27和MGC-803细胞的抗肿瘤作用研究  

Anti-tumor effect of Huaier extract supernatant on human gastric cancer HGC-27 and MGC-803 cells

作　　者：魏雪娇 刘亚鑫 黄惠铭 欧阳里山 谢锦欣 王龙燕 刘东晓 屠鹏飞[1] 胡仲冬 WEI Xue-jiao;LIU Ya-xin;HUANG Hui-ming;OUYANG Li-shan;XIE Jin-xin;WANG Long-yan;LIU Dong-xiao;TU Peng-fei;HU Zhong-dong(Modern Research Center for Traditional Chinese Medicine,School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学中药学院中药现代研究中心,北京100029

出　　处：《中国中药杂志》2022年第23期6457-6465,共9页China Journal of Chinese Materia Medica

基　　金：北京市科技新星计划项目(Z191100001119083)。

摘　　要：探讨槐耳上清(Huaier extract supernatant, HES)对人胃癌HGC-27和MGC-803细胞增殖、凋亡、自噬和体外迁移能力的影响及其分子作用机制。采用HPLC-MS对HES中的主要成分进行简单初步分析。通过MTT法、集落形成实验、EdU染色法探究HES对人胃癌HGC-27和MGC-803细胞增殖的影响。利用Hoechst染色法、流式细胞术明确HES对人胃癌HGC-27和MGC-803细胞凋亡的影响。通过吖啶橙染色法和细胞划痕实验分别研究HES对人胃癌HGC-27和MGC-803细胞自噬和体外迁移能力的影响。借助Western blot检测HES对人胃癌HGC-27和MGC-803细胞中与凋亡、上皮-间质转化(epithelial-mesenchymal transition, EMT)以及信号通路相关的蛋白表达水平的调控作用。结果显示,HES中主要含有一些极性较大的成分。HES能够明显降低人胃癌细胞的细胞活力,且具有浓度和时间依赖性,给药处理人胃癌HGC-27和MGC-803细胞48 h时,其半数抑制浓度(IC_(50))分别为7.56、10.77 g·L^(-1)。同时,HES能够显著抑制人胃癌细胞的集落形成能力和短期增殖能力。8 g·L^(-1)的HES作用于2种人胃癌细胞72 h,细胞凋亡率分别为62.13%±8.92%和54.50%±3.26%。HES还能够促进人胃癌细胞发生自噬,并削弱人胃癌细胞的体外迁移能力。此外,HES能够上调人胃癌细胞中凋亡标志物PARP的剪切水平和上皮细胞标志物E-cadherin的蛋白水平,并下调p-mTOR、p-S6和p-ERK的蛋白水平。因此,HES是槐耳发挥抗肿瘤作用的有效成分之一,其能够抑制人胃癌细胞的增殖和迁移,并诱导其发生凋亡和自噬,而且mTOR信号和ERK信号可能参与了HES的抗胃癌作用。该研究能够为槐耳的深入研究和临床应用提供新的参考,对进一步推动槐耳的科学开发利用具有重要意义。The purpose of this study was to investigate the effect of Huaier extract supernatant(HES) on the proliferation, apoptosis, autophagy, and migration of human gastric cancer HGC-27 and MGC-803 cells and its molecular mechanisms. The main components in HES were preliminarily analyzed by high-performance liquid chromatography-mass spectrometry(HPLC-MS). Methyl thiazolyl tetrazolium(MTT) assay, colony formation assay, and 5-ethynyl-2′-deoxyuridine(EdU) staining assay were used to explore the effect of HES on the proliferation of human gastric cancer HGC-27 and MGC-803 cells. Hoechst staining and flow cytometry assay were used to determine the effect of HES on apoptosis of human gastric cancer HGC-27 and MGC-803 cells. Acridine orange staining and cell scratch assay were used to determine the effect of HES on autophagy and migration of human gastric cancer HGC-27 and MGC-803 cells, respectively. Western blot was used to investigate the regulatory effect of HES on the expression levels of proteins related to apoptosis, epithelial-mesenchymal transition(EMT), and signaling pathways in human gastric cancer HGC-27 and MGC-803 cells. The results showed that HES mainly contained some components with high polarities. HES significantly reduced the cell viability of human gastric cancer cells in a dose-and time-dependent manner. The IC_(50)values after 48 h of HES treatment in human gastric cancer HGC-27 and MGC-803 cells were 7.56 and 10.77 g·L^(-1), respectively. Meanwhile, HES inhibited the colony-forming ability and short-term proliferation of human gastric cancer cells. The apoptosis rates of HGC-27 and MGC-803 cells treated with 8 g·L^(-1)HES for 72 h were 62.13%±8.92% and 54.50%±3.26%, respectively. HES also promoted autophagy in human gastric cancer cells and impaired their migration ability in vitro. Moreover, HES up-regulated the cleavage of the apoptosis marker poly ADP-ribose polymerase(PARP) and the protein expression level of the epithelial cell marker E-cadherin, and down-regulated the protein levels of ph

关 键 词：槐耳上清(HES) 胃癌 增殖 凋亡 mTOR ERK 

分 类 号：R285[医药卫生—中药学]