红景天甙对人胃癌细胞增殖及凋亡影响的研究
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摘要
目的:探讨红景天甙对人胃癌细胞增殖及凋亡的影响及其初步作用机制。方法:不同浓度的红景天甙干预处理胃癌细胞,CCK-8法检测红景天甙对胃癌细胞增殖的影响; Annexin-V/PI流式细胞术检测胃癌细胞凋亡的变化;荧光显微镜观察Hoechst染色后胃癌细胞凋亡形态及核染色质固缩变化情况;流式细胞仪检测红景天甙对胃癌细胞周期的影响; qRT-PCR检测Bcl-2、Bcl-xl、Bax凋亡基因表达; Transwell实验观察胃癌细胞体外侵袭能力的变化。结果:红景天甙显著抑制胃癌细胞增殖,并呈浓度依赖性;明显促进胃癌细胞凋亡发生,引起核染色质固缩,抗凋亡基因Bcl-2、Bcl-xl表达下调,促进凋亡基因Bax表达;阻滞胃癌细胞于G2/M期;明显抑制胃癌细胞的迁徙。结论:红景天甙明显抑制胃癌细胞的增殖并通过Bcl/Bax信号通路诱导其凋亡,提示了中药红景天甙对临床胃癌患者的潜在治疗效应。
Objective: To investigate the effects of salidroside on the proliferation and apoptosis of human gastric cancer cells and it's preliminary mechanism. Methods: After treatment of salidroside with different concentrations on human gastric cancer cells,the effect of salidroside on the proliferation of gastric cancer cells was observed by CCK-8 method. Annexin-V/PI flow cytometry was used to detect the changes of apoptosis of gastric cancer cells. Fluorescence microscopy was used to observe the apoptosis of gastric cancer cells after Hoechst staining. The influence of salidroside on the cell cycle of gastric cancer was detected by flow cytometry. The apoptosis genes of bcl-2,bcl-xl,bax were examinated by qRT-PCR. Transwell experiment was conducted to observe the changes in the invasion ability of gastric cancer cells in vitro. Results: Salidroside dramatically inhibited the proliferation of gastric cancer cells which is dependent of concentration; obviously promoted the apoptosis of gastric cancer cells,caused the phenomenon of chromatin pyknosis,down-regulated the expression of anti-apoptosis gene bcl-2,bcl-xl; up-regulated the expression of apoptosis gene bax; blocked gastric cancer cells in the G2/M period; significantly inhibited the migration of gastric cancer cells. Conclusion: Salidroside obviously inhibits the proliferation of gastric cancer cells and induces apoptosis through Bcl/Bax signaling pathway,which are suggested that salidroside has a potential therapeutic effect on patients with gastric cancer.
Objective: To investigate the effects of salidroside on the proliferation and apoptosis of human gastric cancer cells and it's preliminary mechanism. Methods: After treatment of salidroside with different concentrations on human gastric cancer cells,the effect of salidroside on the proliferation of gastric cancer cells was observed by CCK-8 method. Annexin-V/PI flow cytometry was used to detect the changes of apoptosis of gastric cancer cells. Fluorescence microscopy was used to observe the apoptosis of gastric cancer cells after Hoechst staining. The influence of salidroside on the cell cycle of gastric cancer was detected by flow cytometry. The apoptosis genes of bcl-2,bcl-xl,bax were examinated by qRT-PCR. Transwell experiment was conducted to observe the changes in the invasion ability of gastric cancer cells in vitro. Results: Salidroside dramatically inhibited the proliferation of gastric cancer cells which is dependent of concentration; obviously promoted the apoptosis of gastric cancer cells,caused the phenomenon of chromatin pyknosis,down-regulated the expression of anti-apoptosis gene bcl-2,bcl-xl; up-regulated the expression of apoptosis gene bax; blocked gastric cancer cells in the G2/M period; significantly inhibited the migration of gastric cancer cells. Conclusion: Salidroside obviously inhibits the proliferation of gastric cancer cells and induces apoptosis through Bcl/Bax signaling pathway,which are suggested that salidroside has a potential therapeutic effect on patients with gastric cancer.
引文
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[14]贺拥军,岳秋娟,张敏.藏药红景天甙诱导DC成熟并增强T细胞抑制肺癌的实验研究[J].现代免疫学,2015,35(3):226-229.He YJ,Yue QJ,Zhang M.Experimental study on the induction of DC maturation and enhancement of T cells inhibition of lung cancer by salidroside[J].Modern Immunol,2015,35(3):226-229.
[15]解方为,欧阳学农,蒋明德.红景天甙对人肝癌细胞的诱导分化作用[J].西南国防医药,2005,15(6):613-615.Jie FW,Ouyang XN,Giang MD.The induction and differentiation of salidroside in human hepatocellular carcinoma cells[J].Southwest National Defense Med,2005,15(6):613-615.
[16]Mishra KP,Padwad YS,Dutta A,et al.Aqueous extract of Rhodiola imbricata rhizome inhibits proliferation of an erythroleukemic cell line K-562 by inducing apoptosis and cell cycle arrest at G2/M phase[J].Immunobiology,2008,213(2):125-131.
[17]Hu X,Zhang X,Qiu S,et al.Salidroside induces cell-cycle arrest and apoptosis in human breast cancer cells[J].Biochem Biophys Res Commun,2010,398(1):62-67.
[18]韦丽兰,莫书荣.红景天苷对人胃癌细胞SGC-7901抑瘤作用的体外实验研究[J].山东大学学报(医学版),2015,53(12):12-16.Wei lilan,Mo shurong.In vitro experimental study on the antitumor effect of rhodiodin on human gastric cancer cells sgc-7901[J].J Shandong Univ(medical edition),2015,53(12):12-16.
[2]Chen XZ,Liu Y,Wang R,et al.Improvement of cancer control in mainland China:epidemiological profiles during the 2004-10National Cancer Prevention and Control Program[J].Lancet,2016,388(Suppl 1):S40
[3]Wang R,Zhang MG,Chen XZ,et al.Risk population of Helicobacter pylori infection among Han and Tibetan ethnicities in western China:a cross-sectional,longitudinal epidemiological study[J].Lancet,2016,388(Suppl 1):S17.
[4]Fornaro L,Vasile E,Falcone A.Apatinib in advanced gastric cancer:a doubtful step forward[J].J Clin Oncol,2016,15(8):6931-6933.
[5]Liang L,Ma Z,Dong M,et al.Protective effects of salidroside against isoflurane-induced cognitive impairment in rats[J].Human Exp Toxicol,2017,6(1)DOI:100.1177/0960327116688068.
[6]王林丽.红景天及复方制剂药理和临床研究进展[J].中医药学报,2003,31(1):51-53.Wang LL.Pharmacology and clinical research progress of rhodiola rosea and compound preparation[J].J Traditional Chin Med,2003,31(1):51-53.
[7]Wang S,He H,Chen L,et al.Protective effects of salidroside in the MPTP/MPP(+)-induced model of Parkinson's disease through ROS-NO-related mitochondrion pathway[J].Mol Neurobiol,2015,51(2):718-728.
[8]Luo F,Liu J,Yan T,Miao M:Salidroside alleviates cigarette smoke-induced COPD In mice[J].Biomed Pharmacotherapy,2017,86:155-161.
[9]荣黎、戴立里,曾维政,等.红景天甙对拟高原缺氧大鼠肝损伤的保护作用[J].中国组织工程研究与临床康复,2010,14(31):5813-5817.Rongli,Dai lili,Zeng weizheng,et al.protective effect of rhodiola glycoside on liver injury in hypoxic plateau rats[J].Tissue Engineering Res Clin Rehabilitation China,2010,14(31):5813-5817.
[10]陈亚东,曹秀兰,田长有,等.高山红景天对小鼠耐缺氧、抗疲劳及耐低温作用的影响[J].中国中医药科技,2002,9(3):157-158.Chen yadong,Cao xiulan,Tian changyou,et al.Effect of alpine rhodiola on hypoxia tolerance,fatigue resistance and hypothermia resistance in mice[J].Science Technol Chin Traditional Med,2002,9(3):157-158.
[11]孙非,肖纫霞,孙寒.高山红景天酪醇对病毒性心肌炎小鼠免疫功能及抗氧化酶活性的影响[J].中国药理学通报,2000,16(1):120.Sun fei,Xiao danxia,Sun han.Effect of kaolin on immune function and antioxidant enzyme activity in mice with viral myocarditis[J].Chin Pharmacol Bulletin,2000,16(1):120.
[12]苗艳波,师海波,孙英莲,等.高山红景天总甙的抗衰老作用[J].中药药理与临床,2004,2(5):2-21.Miao yanbo,Shi haibo,Sun yinglian,et al.The anti-aging effect of total glucoside of albino rhodiola[J].Pharmacol Clin Traditional Chin Med,2004,2(5):2-21.
[13]吴晓玲,曾维政,蒋明德,等.红景天甙对肝纤维化大鼠肝组ROCK表达的影响[J].世界华人消化杂志,2009,17(8):765-769.Wu XL,Zeng WZ,Jiang MD,et al.Effect of salidroside on ROCKexpression in hepatic fibrosis rats[J].World Chin Digest,2009,17(8):765-769.
[14]贺拥军,岳秋娟,张敏.藏药红景天甙诱导DC成熟并增强T细胞抑制肺癌的实验研究[J].现代免疫学,2015,35(3):226-229.He YJ,Yue QJ,Zhang M.Experimental study on the induction of DC maturation and enhancement of T cells inhibition of lung cancer by salidroside[J].Modern Immunol,2015,35(3):226-229.
[15]解方为,欧阳学农,蒋明德.红景天甙对人肝癌细胞的诱导分化作用[J].西南国防医药,2005,15(6):613-615.Jie FW,Ouyang XN,Giang MD.The induction and differentiation of salidroside in human hepatocellular carcinoma cells[J].Southwest National Defense Med,2005,15(6):613-615.
[16]Mishra KP,Padwad YS,Dutta A,et al.Aqueous extract of Rhodiola imbricata rhizome inhibits proliferation of an erythroleukemic cell line K-562 by inducing apoptosis and cell cycle arrest at G2/M phase[J].Immunobiology,2008,213(2):125-131.
[17]Hu X,Zhang X,Qiu S,et al.Salidroside induces cell-cycle arrest and apoptosis in human breast cancer cells[J].Biochem Biophys Res Commun,2010,398(1):62-67.
[18]韦丽兰,莫书荣.红景天苷对人胃癌细胞SGC-7901抑瘤作用的体外实验研究[J].山东大学学报(医学版),2015,53(12):12-16.Wei lilan,Mo shurong.In vitro experimental study on the antitumor effect of rhodiodin on human gastric cancer cells sgc-7901[J].J Shandong Univ(medical edition),2015,53(12):12-16.