灵芝孢子油体外抗肿瘤活性比较研究
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摘要
探讨灵芝孢子油对于不同肿瘤细胞的抗肿瘤活性差异,选取人肝癌细胞(Hep G2)、人非小细胞肺癌细胞(A549)、人结肠癌细胞(HCT116)为待检测肿瘤细胞株,采用MTT法进行细胞活力检测,Western blot法检测NF-κB信号通路激活程度,Caspase-3活性检测法检测Caspase-3酶活性来对灵芝孢子油的抗肿瘤活性机制进行初步探讨,并对3种肿瘤细胞的抗肿瘤活性进行比较。经MTT细胞活力实验验证,发现灵芝孢子油对3种肿瘤细胞的生长均具有一定抑制作用,其抑制强度依次为A549细胞、Hep G2细胞、HCT116细胞; Western blot检测结果显示,灵芝孢子油能促进NF-κB通路的激活,3种肿瘤细胞比较下,A549细胞NF-κB信号通路激活程度最强,其次为Hep G2细胞,对HCT116细胞作用最弱; Caspase-3活性检测结果显示,灵芝孢子油能激活Caspase-3依赖的凋亡相关途径,其中对A549细胞Caspase-3酶激活程度最强,对Hep G2细胞作用较强,对HCT116细胞作用最弱。因此,本研究发现,灵芝孢子油对3种肿瘤细胞的生长均具有一定抑制作用,其抗肿瘤活性呈时间剂量依赖性,其机制可能与激活NF-κB通路与Caspase-3凋亡途径加速肿瘤细胞凋亡坏死相关。3种肿瘤细胞生长抑制实验结果显示,灵芝孢子油对A549细胞的抗肿瘤活性最明显,其次为Hep G2细胞,对HCT116细胞的抗肿瘤活性最弱。
To investigate the antitumor activity induced by Ganoderma lucidum spore oil on different tumor cells. Human hepatoma cells( HepG2),human non-small cells lung cancer cells( A549) and human colon cancer cells( HCT116) were selected and tested. Cell viability was determined by MTT assay. Western blot analysis was performed to measure the expression of NF-κB and Caspase-3 activity in order to elucidate the mechanism of apoptotic activity caused by Ganoderma lucidum spore oil and to screen out the most sensitive cancer cell lines to Ganoderma lucidum spore oil. MTT assay demonstrated that Ganoderma lucidum spore oil had a strong inhibitory effect on the growth of three cancer cell lines. Among these cells,A549 cells were most sensitive to Ganoderma lucidum spore oil,followed by HepG2 cells and then by HCT116 cells. The results of Western blot showed that Ganoderma lucidum spore oil could promote the activation of NF-κB pathway,and that the activation of NF-κB signaling pathway in cancer cells treated by Ganoderma lucidum spore oil was stronger in A549 cells, HepG2 cells,HCT116 cells respectively. The detection of Caspase-3 activity showed that ganoderma spore oil could activate Caspase-3 dependent apoptosis pathways,which was more important in A549 cells,HepG2 cells and HCT116 cells. This study found that Ganoderma lucidum spore oil had inhibitory effects on A549, HepG2 and HCT116 cells growth and that its antitumor activity was in time-dose dependence. The mechanism may be related to the activation of NF-κB pathway and the Caspase-3 apoptotic pathway,which could accelerate apoptosis and necrosis of tumor cells. Among the three kinds of cancer cells,A549 cells was most sensitive to Ganoderma lucidum spore oil,followed by the HepG2 cells,and then by HCT116 cells.
To investigate the antitumor activity induced by Ganoderma lucidum spore oil on different tumor cells. Human hepatoma cells( HepG2),human non-small cells lung cancer cells( A549) and human colon cancer cells( HCT116) were selected and tested. Cell viability was determined by MTT assay. Western blot analysis was performed to measure the expression of NF-κB and Caspase-3 activity in order to elucidate the mechanism of apoptotic activity caused by Ganoderma lucidum spore oil and to screen out the most sensitive cancer cell lines to Ganoderma lucidum spore oil. MTT assay demonstrated that Ganoderma lucidum spore oil had a strong inhibitory effect on the growth of three cancer cell lines. Among these cells,A549 cells were most sensitive to Ganoderma lucidum spore oil,followed by HepG2 cells and then by HCT116 cells. The results of Western blot showed that Ganoderma lucidum spore oil could promote the activation of NF-κB pathway,and that the activation of NF-κB signaling pathway in cancer cells treated by Ganoderma lucidum spore oil was stronger in A549 cells, HepG2 cells,HCT116 cells respectively. The detection of Caspase-3 activity showed that ganoderma spore oil could activate Caspase-3 dependent apoptosis pathways,which was more important in A549 cells,HepG2 cells and HCT116 cells. This study found that Ganoderma lucidum spore oil had inhibitory effects on A549, HepG2 and HCT116 cells growth and that its antitumor activity was in time-dose dependence. The mechanism may be related to the activation of NF-κB pathway and the Caspase-3 apoptotic pathway,which could accelerate apoptosis and necrosis of tumor cells. Among the three kinds of cancer cells,A549 cells was most sensitive to Ganoderma lucidum spore oil,followed by the HepG2 cells,and then by HCT116 cells.
引文
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[19] Lin J,Zhang Y,Wang H,et al. Genetic polymorphisms in theapoptosis-associated gene casp3 and the risk of lung cancer inchinese population[J]. PLoS One,2016,11(10):4016-4158.
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[3] Hajjaj H,Mace C,Roberts M,et al. Effect of 26-oxygenosterolsfrom Ganoderma lucidum and their activity as cholesterol synthe-sis inhibitors[J]. Appl Environ Microbiol,2005,71(7):3653-3678.
[4] Suarez-Arroyo IJ,Rosario-Acevedo R,Aguilar-Perez A,et al.Anti-tumor effects of Ganoderma lucidum(reishi)in inflammatorybreast cancer in in vivo and in vitro models[J]. PLoS One,2013,8(2):457-481.
[5] Yuen J,Gohel M. Anticancer effects of Ganoderma lucidum:areview of scientific evidence[J]. Nutr Cancer,2005,53(1):11-17.
[6] Yue G,Chan B,Han X,et al. Immunomodulatory activities ofGanoderma sinense polysaccharides in human immune cells[J].Nutr Cancer,2013,65(5):765-774.
[7] Akinyemiju T,Abera S,Ahmed M,et al. The burden of primaryliver cancer and underlying etiologies from 1990 to 2015 at theglobal,regional,and national level:results from the global burdenof disease study 2015[J]. JAMA Oncol,2017,3(12):1683-1691.
[8] Vilmar A,Sorensen J. Customising chemotherapy in advancednonsmall cell lung cancer:daily practice and perspectives[J].Eur Respir Rev,2011,20(119):45-52.
[9] Testa U,Pelosi E,Castelli G. Colorectal cancer:genetic abnor-malities,tumor progression,tumor heterogeneity,clonal evolutionand tumor-initiating cells[J]. Med Sci(Basel),2018,6(2):336-351.
[10] Qu D,He J,Liu C,et al. Triterpene-loaded microemulsion usingCoix lacryma-jobi seed extract as oil phase for enhanced antitu-mor efficacy:preparation and in vivo evaluation[J]. Int J Nano-medicine,2014,9(5):109-119.
[11] Ben-Neriah Y,Karin M. Inflammation meets cancer,with NF-kappaB as the matchmaker[J]. Nat Immunol,2011,12(8):715-723.
[12] Disis M. Immune regulation of cancer[J]. J Clin Oncol,2010,28(29):4531-4538.
[13] Smyth M,Dunn G,Schreiber R. Cancer immunosurveillance andimmunoediting:the roles of immunity in suppressing tumor devel-opment and shaping tumor immunogenicity[J]. Adv Immunol,2006,90(4):1-50.
[14] Liu Y,Zhou G,Wang Z,et al. NF-kappaB signaling is essentialfor resistance to heat stress-induced early stage apoptosis inhuman umbilical vein endothelial cells[J]. Sci Rep,2015,5(2):135-147.
[15] Nicholson D,Thornberry N. Caspases:killer proteases[J]. TrendsBiochem Sci,1997,22(8):299-306.
[16] Porter A,Ng P,Janicke R. Death substrates come alive[J].Bioessays,1997,19(6):501-507.
[17] Devarajan E,Sahin A,Chen J,et al. Down-regulation of caspase 3in breast cancer:a possible mechanism for chemoresistance[J].Oncogene,2002,21(57):8843-8851.
[18] Xue Y,Wu L,Liu Y,et al. ENTPD5 induces apoptosis in lungcancer cells via regulating caspase 3 expression[J]. PLoS One,2015,10(3):2012-2046.
[19] Lin J,Zhang Y,Wang H,et al. Genetic polymorphisms in theapoptosis-associated gene casp3 and the risk of lung cancer inchinese population[J]. PLoS One,2016,11(10):4016-4158.