银胶菊内酯对人皮肤癌细胞HaCaT和A375凋亡机制的研究
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摘要
目的:研究银胶菊内酯对人皮肤癌细胞HaCaT和A375凋亡的影响。方法:XTT法检测银胶菊内酯对HaCaT和A375细胞的增殖抑制作用;荧光染色法检测细胞形态学变化;ELISA法检测细胞DNA断裂影响;流式细胞仪检测细胞周期。结果:银胶菊内酯明显抑制HaCaT和A375细胞的增殖,IC50值分别为1.45μmol·L-1和2.9μmol·L-1;银胶菊内酯处理过的细胞中出现核碎裂现象,而在对照组细胞中则无此现象;银胶菊内酯诱导HaCaT和A375细胞DNA断裂的长度分别为1.41nm和0.38 nm,当用银胶菊内酯对HaCaT细胞处理24 h后,S期的细胞比例明显增加(P<0.05);当用银胶菊内酯对A375细胞处理24 h后,G0~G1期的细胞比例明显增加(P<0.05)。结论:低浓度的银胶菊内酯能通过分别阻滞HaCaT和A375细胞的S期和G0~G1期诱导细胞凋亡。
OBJECTIVE To study the apoptosis effect of parthenolide on human skin cancer cell HaCaT and A375.METHODS XTT method was used to detect the proliferation inhibiting effects of parthenolide on HaCaT and A375 cells.Fluorescence was used to observe morphological changes of cells.ELISA method was used to detect the DNA fault of cells.Cell cycle was analyzed by flow cytometry.RESULTS Parthenolide significantly inhibited the proliferation of HaCaT and A375 cells,and IC50 values were 1.45μmol·L~(-1) and 2.9μmol·L~(-1).Nuclear fragmentation was observed in parthenolide treated cells,but there was no such phenomenon in the control group cells.Lengths of HaCaT and A375 DNA fractures were 1.41 nm and 0.38 nm.When the HaCaT cells were treated by parthenolide for 24 hours,the percentage of S phase cells increased significantly(P<0.05).When the A375 cells were treated by parthenolide for 24 hours,the percentage of G0-G1 phase cells increased significantly(P<0.05).CONCLUSION Low concentration of parthenolide can induce HaCaT and A375 cell apoptosis through blocking S and G0-G1 phases,respectively.
OBJECTIVE To study the apoptosis effect of parthenolide on human skin cancer cell HaCaT and A375.METHODS XTT method was used to detect the proliferation inhibiting effects of parthenolide on HaCaT and A375 cells.Fluorescence was used to observe morphological changes of cells.ELISA method was used to detect the DNA fault of cells.Cell cycle was analyzed by flow cytometry.RESULTS Parthenolide significantly inhibited the proliferation of HaCaT and A375 cells,and IC50 values were 1.45μmol·L~(-1) and 2.9μmol·L~(-1).Nuclear fragmentation was observed in parthenolide treated cells,but there was no such phenomenon in the control group cells.Lengths of HaCaT and A375 DNA fractures were 1.41 nm and 0.38 nm.When the HaCaT cells were treated by parthenolide for 24 hours,the percentage of S phase cells increased significantly(P<0.05).When the A375 cells were treated by parthenolide for 24 hours,the percentage of G0-G1 phase cells increased significantly(P<0.05).CONCLUSION Low concentration of parthenolide can induce HaCaT and A375 cell apoptosis through blocking S and G0-G1 phases,respectively.
引文
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[13]陈启建,欧阳明安,谢联辉,等.银胶菊(Parthenium hysterophorus)中抗TMV活性成分的分离及活性测定[J].激光生物学报,2008,17(4):544-548.
[14]Ghantous A,Saikali M,Rau T,et al.Inhibition of tumor promotion by parthenolide:epigenetic modulation of p21[J].Cancer Prev Res(Phila),2012,5(11):1298-1309.
[15]徐惠君,贾勇圣,李帅,等.Kuppel因子4调节人乳腺癌细胞对小白菊内酯敏感性的影响[J].中华实验外科杂志,2013,30(10):2224.
[2]钟东明,朱如芳,钟茂耀.银胶菊内酯对PGCL3细胞uPA活性和表达的影响[J].中药材,2005,28(9):800-802.
[3]周敏.银胶菊内酯药理作用的研究进展[J].中国新药杂志,2010,19(14):1225-1228.
[4]Koprowska K,Hartman ML,Sztiller-Sikorska M,et al.Parthenolide enhances dacarbazine activity against melanoma cells[J].Anticancer Drugs,2013,24(8):835-845.
[5]Weislow OS,Kiser R,Fine DL,et al.New soluble-formazan assay for HIV-1 cytopathic effects:application to high-flux screening of synthetic and natural products for AIDS-antiviral activity[J].J Natl Cancer Inst,1989,81(8):577-586.
[6]George VC,Kumar DRN,Suresh PK,et al.A review on the therapeutic potential of parthenolide:a sesquiterpene lactone[J].Int Res J Pharmacol,2012,3(2):69-73.
[7]Ross J,Arnason T,Birnboim C.Low concentrations of the feverfew component parthenolide inhibit in vitro growth of tumor lines in a cytostatic fashion[J].Planta Med,1999,65(2):126-129.
[8]Amorim MH.Gil da costa RM,Lopes C,et al.sesquiterpene lactones:adverse health effects and toxicity mechanisms[J].Crit Rev Toxicol,2013,43(7):559-579.
[9]曹波,牛聪,卢涛,等.五味子乙素对UVB辐射诱导HaCat细胞凋亡的抑制作用及其机制[J].中国药理学通报,2014,30(4):523-527.
[10]肖敏,徐洪来,周薇,等.顺铂对皮肤癌细胞毒性作用的实验研究[J].广西医学,2012,34(10):1315-1317.
[11]高默杰,徐忠伟,王凤梅,等.钠钾泵抑制剂通过调节细胞周期相关蛋白的生成介导肝癌HepG2细胞周期S期阻滞与凋亡[J].中国药理学通报,2010,26(4):452-456.
[12]Cheng Guang,Xie Liping.Parthenolide induces apoptosis and cell cycle arrest of human5637 bladder cancer cells in vitro[J].Molecules,2011,16(8):6758-6768.
[13]陈启建,欧阳明安,谢联辉,等.银胶菊(Parthenium hysterophorus)中抗TMV活性成分的分离及活性测定[J].激光生物学报,2008,17(4):544-548.
[14]Ghantous A,Saikali M,Rau T,et al.Inhibition of tumor promotion by parthenolide:epigenetic modulation of p21[J].Cancer Prev Res(Phila),2012,5(11):1298-1309.
[15]徐惠君,贾勇圣,李帅,等.Kuppel因子4调节人乳腺癌细胞对小白菊内酯敏感性的影响[J].中华实验外科杂志,2013,30(10):2224.