氧化槐果碱对肝癌SMMC-7721细胞增殖与凋亡的影响
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摘要
目的探讨氧化槐果碱对人肝癌SMMC-7721细胞增殖和凋亡的影响,并探讨其可能介导的信号通路。方法将人肝癌SMMC-7721细胞分为对照组、氧化槐果碱处理组(2、4、8、16、32 mmol/L)。将梯次浓度氧化槐果碱作用于肝癌细胞后培养48 h;采用倒置显微镜观察细胞形态变化;采用CCK8法检测不同浓度氧化槐果碱对细胞增殖的影响,采用流式细胞术检测氧化槐果碱对细胞凋亡的影响,通过免疫印迹法检测氧化槐果碱对肝癌细胞B细胞淋巴瘤/白血病-2(Bcl-2),Bcl-2相关X蛋白(Bax)及转录激活因子(STAT)3、STAT5表达水平的影响,分析氧化槐果碱对肝癌SMMC-7721细胞可能的作用。结果氧化槐果碱呈剂量依赖性抑制肝癌SMMC-7721细胞的增殖,并且诱导细胞凋亡。并呈剂量依赖性上调促凋亡Bax的表达,下调抗凋亡蛋白Bcl-2、STAT3、STAT5的表达。结论氧化槐果碱对人肝癌SMMC-7721细胞有抑制增殖、诱导凋亡的作用,可能是通过酪氨酸激酶(JAK)/STAT信号通路下调STAT3、STAT5蛋白激酶表达,从而上调促凋亡Bax基因表达,下调抗凋亡Bcl-2基因表达。
Objective To investigate the effects of oxysophocarpine on the proliferation and apoptosis of human hepatocellular carcinoma SMMC-7721 cells and its possible signaling pathways.Methods Human hepatocellular carcinoma SMMC-7721 cells were divided into control group and oxysophocarpine treatment group(2,4,8,16,32 mmol/L).The effects of different concentrations of oxysophocarpine on cell proliferation were detected by CCK8 method,the effects of oxysophocarpine on cell apoptosis were detected by flow cytometry,and the apoptotic proteins Bax,Bcl-2 were detected by immunoblotting.The effects of signal transduction and transcription activator(STAT) 3 and STAT5 on the expression level were studied.The possible signaling pathway of sophocarpine oxide on SMMC-7721 cells was analyzed.Results Oxysophocarpine inhibits the proliferation of SMMC-7721 cells in a dose-dependent manner and induces apoptosis.The expression of apoptotic Bax was up-regulated in a dose-dependent manner,and the expression of anti-apoptotic proteins Bcl-2,STAT3 and STAT5 was down-regulated.Conclusion Oxysophocarpine can inhibit proliferation and induce apoptosis of human hepatocellular carcinoma SMMC-7721 cells.This may be related to the down-regulation of STAT3 and STAT5 protein kinase expression by tyrosine kinase(JAK)/STAT signaling pathway,thus up-regulation of Bax gene expression and down-regulation of Bcl-2 gene expression.
Objective To investigate the effects of oxysophocarpine on the proliferation and apoptosis of human hepatocellular carcinoma SMMC-7721 cells and its possible signaling pathways.Methods Human hepatocellular carcinoma SMMC-7721 cells were divided into control group and oxysophocarpine treatment group(2,4,8,16,32 mmol/L).The effects of different concentrations of oxysophocarpine on cell proliferation were detected by CCK8 method,the effects of oxysophocarpine on cell apoptosis were detected by flow cytometry,and the apoptotic proteins Bax,Bcl-2 were detected by immunoblotting.The effects of signal transduction and transcription activator(STAT) 3 and STAT5 on the expression level were studied.The possible signaling pathway of sophocarpine oxide on SMMC-7721 cells was analyzed.Results Oxysophocarpine inhibits the proliferation of SMMC-7721 cells in a dose-dependent manner and induces apoptosis.The expression of apoptotic Bax was up-regulated in a dose-dependent manner,and the expression of anti-apoptotic proteins Bcl-2,STAT3 and STAT5 was down-regulated.Conclusion Oxysophocarpine can inhibit proliferation and induce apoptosis of human hepatocellular carcinoma SMMC-7721 cells.This may be related to the down-regulation of STAT3 and STAT5 protein kinase expression by tyrosine kinase(JAK)/STAT signaling pathway,thus up-regulation of Bax gene expression and down-regulation of Bcl-2 gene expression.
引文
[1] 廖春燕,梁健,杨燕,等.苦豆子的药理及应用概述[J].中国民族民间医药,2009,18(5):6-8.
[2] 资源,赵文键,程庆,等.氧化槐果碱对人乳腺癌细胞增殖及凋亡的影响[J].中国临床药理学杂志,2016,32(2):174-177.
[3] 张绪慧,梁磊,蔡长青,等.氧化槐果碱对人结肠腺癌细胞增殖及凋亡的影响[J].广东医学,2015,36(5):660-664.
[4] 吴璟,杨文,郑婕,等.氧化槐果碱对小鼠炎症性疼痛的镇痛作用研究[J].中国比较医学杂志,2017,27(1):54-58.
[5] Liu Y,Xu Y,Ji WD,et al.Anti-tumor activities of matrine and oxymatrine:literature review[J].Tumour Biol,2014,35(6):5111-5119.
[6] 钱利强,沈根海,刘岩.苦参碱类生物碱及其衍生物抗肝癌作用研究进展[J].实用肿瘤杂志,2015,30(1):97-100.
[7] Darnell JE Jr,Kerr IM,Stark GR.Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins[J].Science,1994,264(5164):1415-1421.
[8] 程初勇,黄照河.JAK/STAT信号通道与相关疾病的研究进展[J].湖北科技学院学报(医学版),2016,30(5):448-451.
[9] Osborn JL,Greer SF.Metastatic melanoma cells evade immune detection by silencing STAT1[J].Int J Mol Sci,2015,16(2):4343-4361.
[10] Uchino Y,Iriyama N,Hatta Y,et al.Granulocyte colony-stimulating factor potentiates all-trans retinoic acid-induced granulocytic differentiation in acute promyelocytic leukemia cell line HT93A[J].Cancer Cell Int,2015,15:30.
[11] Kang FB,Wang L,Jia HC,et al.B7-H3 promotes aggression and invasion of hepatocellular carcinoma by targeting epithelial-to-mesenchymal transition via JAK2/STAT3/Slug signaling pathway[J].Cancer Cell Int,2015,15:45.
[12] 王庆华,韩玮,陈达飞,等.槐果碱对肝癌细胞SMMC-7721和肝细胞L-O2生长的影响[J].中医药导报,2018,24(17):31-35,49.
[13] 王庆华,韩玮,陈达飞,等.槐果碱对肝癌细胞SMMC-7721生长与侵袭的影响及机制研究[J].中国中西医结合杂志,2019,39(1):67-71.
[2] 资源,赵文键,程庆,等.氧化槐果碱对人乳腺癌细胞增殖及凋亡的影响[J].中国临床药理学杂志,2016,32(2):174-177.
[3] 张绪慧,梁磊,蔡长青,等.氧化槐果碱对人结肠腺癌细胞增殖及凋亡的影响[J].广东医学,2015,36(5):660-664.
[4] 吴璟,杨文,郑婕,等.氧化槐果碱对小鼠炎症性疼痛的镇痛作用研究[J].中国比较医学杂志,2017,27(1):54-58.
[5] Liu Y,Xu Y,Ji WD,et al.Anti-tumor activities of matrine and oxymatrine:literature review[J].Tumour Biol,2014,35(6):5111-5119.
[6] 钱利强,沈根海,刘岩.苦参碱类生物碱及其衍生物抗肝癌作用研究进展[J].实用肿瘤杂志,2015,30(1):97-100.
[7] Darnell JE Jr,Kerr IM,Stark GR.Jak-STAT pathways and transcriptional activation in response to IFNs and other extracellular signaling proteins[J].Science,1994,264(5164):1415-1421.
[8] 程初勇,黄照河.JAK/STAT信号通道与相关疾病的研究进展[J].湖北科技学院学报(医学版),2016,30(5):448-451.
[9] Osborn JL,Greer SF.Metastatic melanoma cells evade immune detection by silencing STAT1[J].Int J Mol Sci,2015,16(2):4343-4361.
[10] Uchino Y,Iriyama N,Hatta Y,et al.Granulocyte colony-stimulating factor potentiates all-trans retinoic acid-induced granulocytic differentiation in acute promyelocytic leukemia cell line HT93A[J].Cancer Cell Int,2015,15:30.
[11] Kang FB,Wang L,Jia HC,et al.B7-H3 promotes aggression and invasion of hepatocellular carcinoma by targeting epithelial-to-mesenchymal transition via JAK2/STAT3/Slug signaling pathway[J].Cancer Cell Int,2015,15:45.
[12] 王庆华,韩玮,陈达飞,等.槐果碱对肝癌细胞SMMC-7721和肝细胞L-O2生长的影响[J].中医药导报,2018,24(17):31-35,49.
[13] 王庆华,韩玮,陈达飞,等.槐果碱对肝癌细胞SMMC-7721生长与侵袭的影响及机制研究[J].中国中西医结合杂志,2019,39(1):67-71.