柳珊瑚来源的愈创木薁倍半萜生物碱化合物Muriceidine A体外抗肿瘤活性的研究
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摘要
目的对从高领类尖柳珊瑚Muriceides collaris中分离得到的1种愈创木薁倍半萜生物碱Muriceidine A进行体外抗肿瘤活性研究。方法 MTT法测定Muriceidine A对9种人癌细胞系的增殖抑制作用。流式细胞仪检测Muriceidine A对HeLa细胞周期及凋亡的影响。Western Blot检测Muriceidine A对HeLa细胞中周期相关蛋白、凋亡相关蛋白、E6/E7蛋白及其下游信号分子的影响。结果 Muriceidine A对受试9种人癌细胞均显示出较强的抑制活性,且对其中人宫颈腺癌HeLa细胞的抑制作用最强,IC50为17.40μmol·L~(-1)。流式细胞仪分析及Western Blot检测显示,Muriceidine A能够以剂量依赖性的方式使HeLa细胞阻滞在G2/M期,可通过激活Caspase依赖的线粒体凋亡途径诱导HeLa细胞凋亡。进一步研究表明,Muriceidine A能够通过降低HeLa细胞中E6/E7的水平,抑制ERK和STAT3的磷酸化。结论 Muriceidine A体外对人宫颈腺癌细胞HeLa具有较强的细胞毒作用。研究结果为后续对Muriceidine A的深入研究提供了有益的抗肿瘤相关生物学实验依据。
Objective To investigate the in vitro antitumor activity of a novel compound Muriceidine A,which is a guaiazulene sesquiterpene alkaloid reported as a new compound from the gorgonian Muriceides collaris.Methods MTT assay was used to determine the proliferation inhibition effect of Muriceidine A on nine human cancer cell lines.Flow cytometry was used to detect the effect of Muriceidine A on HeLa cell cycle and apoptosis.The effects of Muriceidine A on cycle-associated proteins,apoptosis-related proteins,E6/E7 proteins and downstream signaling molecules in HeLa cells were assessed by Western Blot.Results Muriceidine A could inhibit the cell proliferation of all of the tested nine human cancer cell lines,to a certain extent,with the strongest effect on the human cervical cancer HeLa cells,with an IC50 of 17.40μmol/L.Flow cytometry analysis and Western Blot assay showed that Muriceidine A could arrest HeLa cells in G2/M phase in a dose-dependent manner,and induce HeLa cell apoptosis via activating the Caspase-dependent mitochondrial apoptosis pathway.Further studies had showed that Muriceidine A inhibited the phosphorylation of ERK and STAT3 via down-regulating the E6/E7 expression level in HeLa cells.Conclusion Muriceidine A had a strong cytotoxic effect on human cervical cancer cell line HeLa in vitro.The present result provided useful information relating to the antitumor property of muriceidine A for the future studies on the muriceidine A and related compounds.
Objective To investigate the in vitro antitumor activity of a novel compound Muriceidine A,which is a guaiazulene sesquiterpene alkaloid reported as a new compound from the gorgonian Muriceides collaris.Methods MTT assay was used to determine the proliferation inhibition effect of Muriceidine A on nine human cancer cell lines.Flow cytometry was used to detect the effect of Muriceidine A on HeLa cell cycle and apoptosis.The effects of Muriceidine A on cycle-associated proteins,apoptosis-related proteins,E6/E7 proteins and downstream signaling molecules in HeLa cells were assessed by Western Blot.Results Muriceidine A could inhibit the cell proliferation of all of the tested nine human cancer cell lines,to a certain extent,with the strongest effect on the human cervical cancer HeLa cells,with an IC50 of 17.40μmol/L.Flow cytometry analysis and Western Blot assay showed that Muriceidine A could arrest HeLa cells in G2/M phase in a dose-dependent manner,and induce HeLa cell apoptosis via activating the Caspase-dependent mitochondrial apoptosis pathway.Further studies had showed that Muriceidine A inhibited the phosphorylation of ERK and STAT3 via down-regulating the E6/E7 expression level in HeLa cells.Conclusion Muriceidine A had a strong cytotoxic effect on human cervical cancer cell line HeLa in vitro.The present result provided useful information relating to the antitumor property of muriceidine A for the future studies on the muriceidine A and related compounds.
引文
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[2]Li P,Liu X,Zhu H Y,et al.Unusual inner-salt guaiazulene alkaloids and bis-sesquiterpene from the South China Sea gorgonian Muriceides Collaris[J].Sci Rep,2017,7(1):7697-7705.
[3]Wallace N,Robinson K,Galloway D A.Beta human papillomavirus E6expression inhibits stabilization of p53and increases tolerance of genomic instability[J].J Viro,2014,88(11):6112-6127.
[4]Zhang W,Che Q,Tan H,et al.Marine Streptomyces sp.derived antimycin analogues suppress HeLa cells via depletion HPV E6/E7mediated by ROS-dependent ubiquitin-proteasome system[J].Sci Rep,2017,7(8):42180-42193.
[5]Gavet O,Pines J.Progressive activation of cyclin B1-Cdk1coordinates entry to mitosis[J].Dev Cell,2010,18(4):533-543.
[6]Ujiki M,Ding X,Salabat M,et al.Apigenin inhibits pancreatic cancer cell proliferation through G2/M cell cycle arrest[J].Mol Cancer,2006,5(5):76-83.
[7]Alabsi A,Lim K,Paterson I,et al.Cell cycle arrest and apoptosis induction via modulation of mitochondrial integrity by Bcl-2family members and caspase dependence in Dracaena cinnabari-treated H400human oral squamous cell carcinoma[J].Biomed Res Int,2016,2016:4904016-4904029.
[8]Ola M,Nawaz M,Ahsan H.Role of Bcl-2family proteins and caspases in the regulation of apoptosis[J].Mol Cell Biochem,2011,351(1/2):41-58.
[9]Li C,Fan S,Owonikoko T,et al.Oncogenic role of EAPII in lung cancer development and its activation of the MAPKERK pathway[J].Oncogene,2011,30(35):3802-3812.
[10]Shukla S,Mahata S,Shishodia G,et al.Functional regulatory role of Stat3in HPV16-mediated cervical carcinogenesis[J].PloS One,2013,8(7):67849-67861.
[11]Chen C,Hsieh F,Lieblein J,et al.Stat3activation in human endometrial and cervical cancers[J].Bri J Cancer,2007,96(4):591-599.
[12]Zhou P,Weghorst C M,Weinstein I B.Overexpression of cyclin D1 enhances gene amplification[J].Cancer Research,1996,56(1):36-39.
[13]Altieri D C.Survivin,versatile modulation of cell division and apoptosis in cancer[J].Oncogene,2003,22(53):8581-8589.
[14]Vazhappilly C G,Saleh E,Ramadan W,et al.Inhibition of SHP2by new compounds induces differential effects on RAS/RAF/ERK and PI3K/AKT pathways in different cancer cell types[J].Inv New Drugs,2018,1(10):1-10.
[15]Li X,Lu Y,Liang K,et al.Differential responses to doxorubicin-induced phosphorylation and activation of Akt in human breast cancer cells[J].Breast Cancer Res,2005,7(5):589-597.