和厚朴酚促进组织因子途径抑制物-2表达并诱导神经胶质瘤细胞凋亡
|
摘要
目的探究和厚朴酚对神经胶质瘤细胞凋亡的作用及组织因子途径抑制剂因子TFPI-2在其中的分子机制。方法在体外培养神经胶质瘤细胞系U87b,使用不同浓度(0、40、80μM)的和厚朴酚处理神经胶质瘤细胞,利用流式细胞术检测细胞凋亡情况,利用western blot检测TFPI-2、BAX、BCL-2、Cleaved caspase3的蛋白表达情况。利用siRNA沉默U87细胞中TFPI-2的表达,进一步确定TFPI-2的作用。结果 40μM和厚朴酚处理诱导的U87细胞凋亡率(26. 3±0. 55)%明显高于对照组(4. 8±0. 22)%,P <0. 001);相比于对照组(4. 8±0. 22)%,80μM和厚朴酚[(79. 1±0. 31)%,P <0. 001]同样也显著增加细胞凋亡;和厚朴酚成浓度依赖性增加U87细胞内凋亡相关基因BAX/BCL-2比值、Cleaved caspase3的表达以及TFPI-2的蛋白表达;和厚朴酚+si TFP1-2组的细胞凋亡率(12. 7±0. 29)%明显低于和厚朴酚+NC组细胞凋亡率[(57. 2±0. 55)%,P <0. 001];TFPI-2沉默能明显抑制促凋亡蛋白BAX、Cleaved caspase3的表达水平,并促进抑凋亡蛋白BCL-2的表达。结论和厚朴酚可以通过诱导U87神经胶质瘤细胞中TFPI-2高表达,进而促进细胞凋亡的发生。
Objective To explore the effect of honokiol on glioma cell and its cellular mechanism with tissue factor pathway inhibitor-2( TFPI-2) involved. Methods The glioma cell line U87 b was cultured in vitro,with honokiol-treated glioma cells,at different concentrations( 0,40,80 μM),using flow cytometry,the cell apoptosis was detected. Using western blotting test,the protein expressions of TFPI-2,BAX,BCL02 and cleaved caspase-3 were determined. Using siRNA,the expression of TFPI-2 in U87 cells was silenced. Results Induced by honokion,at the concentration of 40μM,U87 apoptosis( 26. 3 ± 0. 55) % was higher obviously than the control group( 4. 8 ± 0. 22) %,P < 0. 001. Compared with the control group( 4. 8 ± 0. 22) %,the apoptosis with the induction of 80μM honokiol( 79. 1 ± 0. 31) was increased significantly( P < 0. 001). BAX/BCL-2 ratio,cleaved caspase-3 expression and TFPI-2 expression were increased in dependence with the concentrations of honokiol. The apoptosis in the group of honokiol + si TFPI-2( 12. 7 ± 0. 29) % was lower obviously than the group of honokiol and NC( 57. 2 ± 0. 55) %,P < 0. 001. TFPI-2 silencing obviously inhibited the expressions of proapoptotic protein BAX and cleaved caspase-3,promoted the expression of the inhibitory protein BCL-2. Conclusion Honokiol promotes cell apoptosis through inducing TFPI-2 expression in U87 glioma cells.
Objective To explore the effect of honokiol on glioma cell and its cellular mechanism with tissue factor pathway inhibitor-2( TFPI-2) involved. Methods The glioma cell line U87 b was cultured in vitro,with honokiol-treated glioma cells,at different concentrations( 0,40,80 μM),using flow cytometry,the cell apoptosis was detected. Using western blotting test,the protein expressions of TFPI-2,BAX,BCL02 and cleaved caspase-3 were determined. Using siRNA,the expression of TFPI-2 in U87 cells was silenced. Results Induced by honokion,at the concentration of 40μM,U87 apoptosis( 26. 3 ± 0. 55) % was higher obviously than the control group( 4. 8 ± 0. 22) %,P < 0. 001. Compared with the control group( 4. 8 ± 0. 22) %,the apoptosis with the induction of 80μM honokiol( 79. 1 ± 0. 31) was increased significantly( P < 0. 001). BAX/BCL-2 ratio,cleaved caspase-3 expression and TFPI-2 expression were increased in dependence with the concentrations of honokiol. The apoptosis in the group of honokiol + si TFPI-2( 12. 7 ± 0. 29) % was lower obviously than the group of honokiol and NC( 57. 2 ± 0. 55) %,P < 0. 001. TFPI-2 silencing obviously inhibited the expressions of proapoptotic protein BAX and cleaved caspase-3,promoted the expression of the inhibitory protein BCL-2. Conclusion Honokiol promotes cell apoptosis through inducing TFPI-2 expression in U87 glioma cells.
引文
[1]Desjardins A,Gromeier M,Herndon JE,et al. Recurrent Glioblastoma Treated with Recombinant Poliovirus[J]. N Engl J Med,2018,379(2):150-161.
[2]Touat M,Idbaih A,Sanson M,et al. Glioblastoma targeted therapy:updated approaches from recent biological insights[J]. Annals of oncology:official journal of the European Society for Medical Oncology,2017,28(7):1457-1472.
[3]Wu G,Wang W,Liu Y,et al. NY-SAR-35 is involved in apoptosis,cell migration,invasion and epithelial to mesenchymal transition in glioma[J]. Biomed Pharmacother,2018(97):1632-1638.
[4]Xu ZF,Sun XK,Chen G,et al. Oroxyloside inhibits human glioma progression by suppressing proliferation,metastasis and inducing apoptosis related pathways[J]. Biomed Pharmacother,2018(97):1564-1574.
[5]Zhang J,Liu Q,Shi L,et al. Honokiol triggers receptor interacting protein kinase 3 mediated cell death of neuroblastoma cells by upregulating reactive oxygen species[J]. Mol Med Rep,2017,16(6):8525-8529.
[6]Balan M,Chakraborty S,Flynn E,et al. Honokiol inhibits c-MetHO-1 tumor-promoting pathway and its cross-talk with calcineurin inhibitor-mediated renal cancer growth[J]. Sci Rep,2017,7(1):5900.
[7]Zhang M,Yue B,Zhang AH,et al. TC38,a teleost TFPI-2 peptide that kills bacteria via penetration of the cell membrane and interaction with nucleic acids[J]. Fish Shellfish Immunol,2017(64):104-110.
[8]Wang G,Zeng Y,Chen S,et al. Localization of TFPI-2 in the nucleus modulates MMP-2 gene expression in breast cancer cells[J].Scientific reports,2017,7(1):13575.
[9]魏熙胤,臧凤琳,孙保存. TFPI-2对恶性肿瘤细胞增殖凋亡侵袭转移调控作用的研究进展[J].中国肿瘤临床,2014(21):1399-1402.
[10]Li Z,Xu Y,Wang Q,et al. Tissue factor pathway inhibitor-2 induced hepatocellular carcinoma cell differentiation[J]. Saudi journal of biological sciences,2017,24(1):95-102.
[11]樊英怡,李倩,梁晨,等.中医药干预乳腺癌肿瘤微环境与血管新生的研究思路与进展[J].世界中西医结合杂志,2013,8(8):856-860.
[12]Feng C,Ho Y,Sun C,et al. TFPI-2 expression is decreased in bladder cancer and is related to apoptosis[J]. Journal of BUON:official journal of the Balkan Union of Oncology,2016,21(6):1518-1523.
[13]Talarek S,Listos J,Barreca D,et al. Neuroprotective effects of honokiol:from chemistry to medicine[J]. BioFactors(Oxford,England),2017,43(6):760-769.
[14]Prasad R,Katiyar SK. Honokiol,an Active Compound of Magnolia Plant,Inhibits Growth,and Progression of Cancers of Different Organs[J]. Advances in experimental medicine and biology,2016(928):245-265.
[15]吴秋玲,孔庆志.中药抗肿瘤血管生成的研究进展[J].世界中西医结合杂志,2010,5(1):87-89.
[2]Touat M,Idbaih A,Sanson M,et al. Glioblastoma targeted therapy:updated approaches from recent biological insights[J]. Annals of oncology:official journal of the European Society for Medical Oncology,2017,28(7):1457-1472.
[3]Wu G,Wang W,Liu Y,et al. NY-SAR-35 is involved in apoptosis,cell migration,invasion and epithelial to mesenchymal transition in glioma[J]. Biomed Pharmacother,2018(97):1632-1638.
[4]Xu ZF,Sun XK,Chen G,et al. Oroxyloside inhibits human glioma progression by suppressing proliferation,metastasis and inducing apoptosis related pathways[J]. Biomed Pharmacother,2018(97):1564-1574.
[5]Zhang J,Liu Q,Shi L,et al. Honokiol triggers receptor interacting protein kinase 3 mediated cell death of neuroblastoma cells by upregulating reactive oxygen species[J]. Mol Med Rep,2017,16(6):8525-8529.
[6]Balan M,Chakraborty S,Flynn E,et al. Honokiol inhibits c-MetHO-1 tumor-promoting pathway and its cross-talk with calcineurin inhibitor-mediated renal cancer growth[J]. Sci Rep,2017,7(1):5900.
[7]Zhang M,Yue B,Zhang AH,et al. TC38,a teleost TFPI-2 peptide that kills bacteria via penetration of the cell membrane and interaction with nucleic acids[J]. Fish Shellfish Immunol,2017(64):104-110.
[8]Wang G,Zeng Y,Chen S,et al. Localization of TFPI-2 in the nucleus modulates MMP-2 gene expression in breast cancer cells[J].Scientific reports,2017,7(1):13575.
[9]魏熙胤,臧凤琳,孙保存. TFPI-2对恶性肿瘤细胞增殖凋亡侵袭转移调控作用的研究进展[J].中国肿瘤临床,2014(21):1399-1402.
[10]Li Z,Xu Y,Wang Q,et al. Tissue factor pathway inhibitor-2 induced hepatocellular carcinoma cell differentiation[J]. Saudi journal of biological sciences,2017,24(1):95-102.
[11]樊英怡,李倩,梁晨,等.中医药干预乳腺癌肿瘤微环境与血管新生的研究思路与进展[J].世界中西医结合杂志,2013,8(8):856-860.
[12]Feng C,Ho Y,Sun C,et al. TFPI-2 expression is decreased in bladder cancer and is related to apoptosis[J]. Journal of BUON:official journal of the Balkan Union of Oncology,2016,21(6):1518-1523.
[13]Talarek S,Listos J,Barreca D,et al. Neuroprotective effects of honokiol:from chemistry to medicine[J]. BioFactors(Oxford,England),2017,43(6):760-769.
[14]Prasad R,Katiyar SK. Honokiol,an Active Compound of Magnolia Plant,Inhibits Growth,and Progression of Cancers of Different Organs[J]. Advances in experimental medicine and biology,2016(928):245-265.
[15]吴秋玲,孔庆志.中药抗肿瘤血管生成的研究进展[J].世界中西医结合杂志,2010,5(1):87-89.