蒲公英萜醇对人乳腺癌细胞MCF-7增殖及凋亡的影响
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摘要
目的:探讨蒲公英萜醇对人乳腺癌细胞MCF-7增殖、凋亡的影响及其可能的分子机制。方法:体外培养人乳腺癌细胞MCF-7,采用噻唑蓝比色法和平板克隆实验检测蒲公英萜醇对MCF-7细胞增殖的影响;Hoechst 33342荧光染色法检测细胞凋亡的形态变化;Western blot法检测凋亡相关蛋白的表达。结果:蒲公英萜醇对人乳腺癌细胞MCF-7有明显的增殖抑制作用(P<0.05);Hoechst染色结果显示,给药后细胞核内染色质凝集,核固缩,可见凋亡小体;Western blot结果显示,蒲公英萜醇可极显著提高Bax/Bcl-2的比值(P<0.01)以及上调活化型Caspase-3/9的表达(P<0.01),下调PARP的表达(P<0.01)。结论:蒲公英萜醇能够明显抑制人乳腺癌细胞MCF-7增殖,并通过线粒体途径诱导其凋亡。
Objective: To investigate the effect of taraxerol on the proliferation and apoptosis of breast cancer MCF-7 cells and to explore its possible mechanism of action. Methods: The effect of taraxerol on the proliferation of MCF-7 cells was detected by methylthiazolyldiphenyltetrazolium bromide assay and colony formation assay; cell apoptosis was tested by Hoechst 33342 fluorescence staining, and apoptosis-related protein expression was detected by Western blot. Results: Taraxerol significantly inhibited the proliferation of MCF-7 cells(P < 0.05). Hoechst staining revealed a series of morphological changes in MCF-7 cells associated with apoptosis, and Western blot data indicated that taraxerol significantly increased the ratio of Bax/Bcl-2(P < 0.01) and up-regulated cleaved caspase-3/9 expression(P < 0.01), and down-regulated PARP expression(P < 0.01). Conclusion: Taraxerol can inhibit the proliferation, and induce the apoptosis of MCF-7 cells via the mitochondrial signaling pathway.
Objective: To investigate the effect of taraxerol on the proliferation and apoptosis of breast cancer MCF-7 cells and to explore its possible mechanism of action. Methods: The effect of taraxerol on the proliferation of MCF-7 cells was detected by methylthiazolyldiphenyltetrazolium bromide assay and colony formation assay; cell apoptosis was tested by Hoechst 33342 fluorescence staining, and apoptosis-related protein expression was detected by Western blot. Results: Taraxerol significantly inhibited the proliferation of MCF-7 cells(P < 0.05). Hoechst staining revealed a series of morphological changes in MCF-7 cells associated with apoptosis, and Western blot data indicated that taraxerol significantly increased the ratio of Bax/Bcl-2(P < 0.01) and up-regulated cleaved caspase-3/9 expression(P < 0.01), and down-regulated PARP expression(P < 0.01). Conclusion: Taraxerol can inhibit the proliferation, and induce the apoptosis of MCF-7 cells via the mitochondrial signaling pathway.
引文
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[2]FOUNTZILAS G,DAFNI U,PAPADIMITRIOU C,et al.Dosedense sequential adjuvant chemotherapy followed,as indicated,by trastuzumab for one year in patients with early breast cancer:first report at 5-year median follow-up of a Hellenic Cooperative Oncology Group randomized phaseⅢtrial[J].BMC Cancer,2014,14(1):1-14.DOI:10.1186/1471-2407-14-515.
[3]林云,江林,蒋健,等.蒲公英的药理作用研究进展[J].中国现代中药,2011,13(8):42-47.
[4]杨岚,李华峰,刁海鹏,等.蒲公英花中总酚酸和总黄酮含量测定及其抗氧化性能研究[J].食品科学,2011,32(17):160-163.
[5]李景华,刘玉芹,王黎明.蒲公英属植物研究进展[J].吉林医药学院学报,2011,32(3):160-166.
[6]章雪.蒲公英和甘草代谢物的抗肿瘤活性物质基础研究[D].上海:上海交通大学,2013:74-76.
[7]SHARMA K,ZAFAR R.Occurrence of taraxerol and taraxasterol in medicinal plants[J].Pharmacognosy Review,2015,9(17):19-23.DOI:10.4103/0973-7847.156317.
[8]LI S,SHI Y,SHANG X Y,et al.Triter penoids from the roots of Pterospermum heterophyllum Hance.[J].Journal of Asian Natural Products Research,2009,11(7):652-657.DOI:10.1080/10286020902964248.
[9]谭宝,石海莲,季光,等.蒲公英萜醇和乙酰蒲公英萜醇对胃癌细胞株AGS细胞周期和凋亡的影响[J].中西医结合学报,2011,9(6):638-642.
[10]KAENNAKAM S,SICHAEM J,KHUMKRATOK S,et al.A new taraxerol derivative from the roots of Microcos tomentosa[J].Natural Product Communications,2013,8(10):1371-1372.
[11]齐绪林,高鹏飞,乔田奎,等.中药蒲公英抗肿瘤作用研究进展[J].中国肿瘤,2015,24(1):53-56.
[12]宋长城,朱美玲.中药所含三萜类化合物抗肿瘤活性及其作用机制的研究进展[J].现代肿瘤医学,2011,19(9):1880-1883.
[13]陈红林,乔华,孙体健.蒲公英花提取物的体外抗肿瘤活性研究[J].中国药物与临床,2014,14(9):1179-1181;1309.
[14]孙常松,李玛琳.五环三萜类化合物抗肿瘤活性及其机制研究进展[J].中国民族民间医药,2009,18(12):14-15.
[15]MOSMANN T.Rapid colorimetric assay for cellular growth and survival:application to proliferation and cytotoxicity assays[J].Journal of Immunological Methods,1983,65(1/2):55-63.
[16]苏方,刘浩,程秀,等.塞来昔布联合多柔比星对乳腺癌细胞增殖及凋亡的影响[J].中国药理学通报,2011,27(5):632-638.
[17]BERRIDGE M V,HERST P M,TAN A S.Tetrazolium dyes as tools in cell biology:new insights into their cellular reduction[J].Biotechnology Annual Review,2005,11:127-152.DOI:10.1016/S1387-2656(05)11004-7.
[18]SAVILL J,FADOK V.Corpse clearance defines the meaning of cell death[J].Nature,2000,407:784-788.DOI:10.1038/35037722.
[19]朱彩平,张声华,肖军霞.枸杞多糖诱导人宫颈癌He La细胞凋亡的形态学研究[J].食品科学,2010,31(19):329-334.
[20]YAO X Y,LU B Y,LüC T,et al.Taraxerol induces cell apoptosis through mitochondria-mediated pathway in He La cells[J].Cell Journal,2017,19(3):512-519.DOI:10.22074/cellj.2017.4543.
[21]TISCHNER D,WOESS C,OTTINA E,et al.Bcl2 regulated cell death signalling in the prevention of autoimmunity[J].Cell Death&Disease,2010,3(1):1-11.DOI:10.1038/cddis.2010.27.
[22]KANG M H,REYNOLDS C P.Bcl-2 inhibitors:targeting mitochondrial apoptotic pathways in cancer therapy[J].Clinical Cancer Research,2009,15(4):1126-1132.DOI:10.1158/1078-0432.CCR-08-0144.
[23]李谣,陈金龙,王丽颖,等.羊肚菌多糖抑制人乳腺癌细胞MDAMB-231增殖和诱导细胞凋亡研究[J].食品科学,2016,37(21):214-218.DOI:10.7506/spkx1002-6630-201621036.
[24]高超,华子春.细胞凋亡检测方法新进展[J].中国细胞生物学学报,2011,33(5):564-569.
[25]LEVINE B,SINHA S,KROEMER G.Bcl-2 family members:dual regulators of apoptosis and autophagy[J].Autophagy,2008,4:600-606.DOI:10.4161/auto.6260.
[26]LIU G Y,WANG T,WANG T,et al.Effects of apoptosis-related proteins Caspase-3,Bax and Bcl-2 on cerebral ischemia rats[J].Biomedical Reports,2013,1(6):861-867.DOI:10.3892/br.2013.153.
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[28]CHEN L Y,JIN T F,ZHU K,et al.PI3K/m TOR dual inhibitor BEZ235 and histone deacetylase inhibitor trichostatin A synergistically exert anti-tumor activity in breast cancer[J].Oncotarget,2017,8(7):11937-11949.DOI:10.18632/oncotarget.14442.