血竭石油醚提取物对人胃癌HGC-27和MGC-803细胞的抗肿瘤作用  

Anti-tumor Effect of Draconis Sanguis Petroleum Ether Fraction on Human Gastric Cancer HGC-27 and MGC-803 Cells

作　　者：黄惠铭 田颖颖 庞道然 刘亚鑫 欧阳里山 屠鹏飞[1] 李军[1] 胡仲冬 HUANG Hui-ming;TIAN Ying-ying;PANG Dao-ran;LIUYa-xin;OUYANG Li-shan;TU Peng-fei;LI Jun;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：《中国实验方剂学杂志》2022年第1期85-91,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：北京市科技新星计划项目(Z191100001119083);中华中医药学会青年人才托举工程项目[CACM-2018-(QNRC2-B05)]。

摘　　要：目的:研究血竭石油醚提取物(SDPEF)对人胃癌HGC-27和MGC-803细胞增殖、凋亡、体外迁移能力及自噬的影响,并阐明其分子作用机制。方法:采用细胞增殖与活性检测(CCK-8)法检测SDPEF(0,20,40,60,80 mg·L^(-1))作用24,48,72 h时,对人胃癌HGC-27和MGC-803细胞体外增殖的影响。借助Hoechst染色实验观察不同浓度的SDPEF作用48 h后对HGC-27和MGC-803细胞凋亡的影响。采用流式细胞术检测不同浓度SDPEF作用48 h,对HGC-27和MGC-803细胞凋亡率的影响。采用细胞划痕实验观察不同浓度SDPEF在不同作用时间点对人胃癌HGC-27和MGC-803细胞体外迁移能力的影响。借助吖啶橙染色实验检测不同浓度SDPEF对HGC-27和MGC-803细胞自噬的影响。利用蛋白免疫印迹法检测SDPEF作用人胃癌HGC-27和MGC-803细胞48 h后对信号通路相关蛋白表达水平的调控作用。结果:与空白组比较,SDPEF(30 mg·L^(-1))组人胃癌HGC-27和MGC-803细胞的增殖和体外迁移能力明显降低(P<0.05),且具有浓度与时间依赖性;SDPEF(60 mg·L^(-1))还能够诱导人胃癌HGC-27和MGC-803细胞凋亡(P<0.01)和自噬的发生。与空白组比较,SDPEF(60 mg·L^(-1))能够下调磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)表达(P<0.05,P<0.01),且能够下调磷酸化信号传导与转录激活因子3(p-STAT3)蛋白表达水平(P<0.01),提示SDPEF可能是通过抑制mTOR/STAT3信号通路来抑制人胃癌HGC-27和MGC-803细胞的增殖、迁移并诱导其发生凋亡和自噬。结论:mTOR/STAT3信号通路下调可能参与了SDPEF的抗胃癌作用。该研究能够为血竭抗肿瘤作用研究提供一定的参考。Objective:To investigate the effect of Draconis Sanguis petroleum ether fraction(DSPEF on the proliferation,apoptosis,migration,and autophagy of human gastric cancer HGC-27 and MGC-803 cells,and preliminarily elucidate its molecular mechanism. Method: Cell counting kit-8(CCK-8)assay was used to detect the effect of DSPEF at different concentrations(0,20,40,60,80 mg·L^(-1))on the proliferation of HGC-27 and MGC-803 cells after 24,48,72 h. Hoechst staining and flow cytometry were used to explore the effects of DSPEF at different concentrations on the apoptosis and apoptosis rate of HGC-27 and MGC-803 cells after48 h treatment,respectively. The wound healing assay and acridine orange staining were used to investigate the effects of DSPEF on the migration and autophagy of HGC-27 and MGC-803 cells,respectively. Western blot was used to detect the expression levels of signaling pathway-related proteins in HGC-27 and MGC-803 cells treated with DSPEF for 48 h. Result: Compared with the control group,DSPEF(30 mg·L^(-1))inhibited the proliferation and migration of HGC-27 and MGC-803 cells in a concentration-and time-dependent manner(P<0.05),and induced the apoptosis(P<0.01)and autophagy of HGC-27 and MGC-803 cells. DSPEF(60 mg·L^(-1))down-regulated the protein levels of phosphorylated mammalian target of rapamycin(p-mTOR)(P<0.05,P<0.01)and down-regulated phospho-signal transducer and activator of transcription 3(p-STAT3)in HGC-27 and MGC-803 cells(P<0.01),suggesting that DSPEF presumedly inhibited the proliferation and migration of human gastric cancer HGC-27 and MGC-803 cells and induced their apoptosis and autophagy by inhibiting the mTOR/STAT3 signaling pathway. Conclusion: The down-regulation of the mTOR/STAT3 signaling pathway may be involved in the anti-gastric cancer effect of DSPEF. This study is expected to provide a reference for the investigation of the anti-tumor effect of Draconis Sanguis.

关 键 词：血竭石油醚提取物 人胃癌细胞 增殖 凋亡 迁移 自噬 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]