异甘草素对小儿血管瘤内皮细胞生物学特性及PI3K/AKT信号通路的影响
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摘要
目的:探讨异甘草素对小儿血管瘤内皮细胞生物学特性及PI3K/AKT信号通路的影响。方法:将培养的小儿血管瘤内皮细胞分为4组,分别用0、20、40和80μmol/L异甘草素处理24、48、72 h,采用MTT法检测细胞活力。处理72 h后,采用Transwell小室实验检测细胞的侵袭能力,流式细胞仪检测细胞周期变化和凋亡,实时荧光定量PCR和Western blot检测细胞中Cleaved Caspase-3、Bax、Cyclin B1、MMP-9和p-AKT mRNA和蛋白的表达。结果:异甘草素能够呈时间、浓度依赖性地抑制小儿血管瘤内皮细胞生长;随异甘草素作用浓度的升高,侵袭细胞数和G0/G1期细胞百分比均逐渐降低,而G2/M期细胞百分比及细胞凋亡率均逐渐升高;Cleaved Caspase-3、Bax mRNA和蛋白表达水平逐渐升高,而Cyclin B1、MMP-9、p-AKT mRNA和蛋白表达水平逐渐下降(P<0.05)。结论:异甘草素可能通过调控PI3K/AKT信号通路抑制小儿血管瘤内皮细胞的生长和侵袭,诱导细胞周期阻滞,并促进细胞凋亡。
Aim:To investigate the effects of isoliquiritigenin on biological characteristics and PI3 K/AKT signaling pathway of infantile hemangioma endothelial cells.Methods:The cultured infantile hemangioma endothelial cells were allocated into 4 groups,and were treated by 0,20,40 and 80 μmol/L isoliquiritigenin for 24,48,and 72 hours,and cell viability was detected by MTT method.After 72 hour treatment,the invasion ability of the cells was detected by the Transwell chamber test,and cell cycle changes and apoptosis were measured by flow cytometry.The expressions of Cleaved Caspase-3,Bax,Cyclin B1,MMP-9 and p-AKT mRNA and protein in cells were tested by qRT-PCR and Western blot.Results:Isoliquiritigenin inhibited the growth of the cells in a time-and concentration-dependent manner(P<0.05).With the increase of isoliquiritigenin concentration,the number of invasive cells and the percentage of G0/G1 phase cells decreased gradually,while the percentage of cells in G2/M phases and apoptosis rate of the cells increased gradually(P<0.05);the expressions of Cleaved Caspase-3 and Bax mRNA and protein increased gradually,while the expressions of Cyclin B1,MMP-9,p-AKT mRNA and protein decreased gradually(P<0.05).Conclusion:Isoliquiritigenin may inhibit the growth and invasion of infantile hemangioma endothelial cells,induce cell cycle arrest and promote cell apoptosis by regulating PI3 K/AKT signaling pathway.
Aim:To investigate the effects of isoliquiritigenin on biological characteristics and PI3 K/AKT signaling pathway of infantile hemangioma endothelial cells.Methods:The cultured infantile hemangioma endothelial cells were allocated into 4 groups,and were treated by 0,20,40 and 80 μmol/L isoliquiritigenin for 24,48,and 72 hours,and cell viability was detected by MTT method.After 72 hour treatment,the invasion ability of the cells was detected by the Transwell chamber test,and cell cycle changes and apoptosis were measured by flow cytometry.The expressions of Cleaved Caspase-3,Bax,Cyclin B1,MMP-9 and p-AKT mRNA and protein in cells were tested by qRT-PCR and Western blot.Results:Isoliquiritigenin inhibited the growth of the cells in a time-and concentration-dependent manner(P<0.05).With the increase of isoliquiritigenin concentration,the number of invasive cells and the percentage of G0/G1 phase cells decreased gradually,while the percentage of cells in G2/M phases and apoptosis rate of the cells increased gradually(P<0.05);the expressions of Cleaved Caspase-3 and Bax mRNA and protein increased gradually,while the expressions of Cyclin B1,MMP-9,p-AKT mRNA and protein decreased gradually(P<0.05).Conclusion:Isoliquiritigenin may inhibit the growth and invasion of infantile hemangioma endothelial cells,induce cell cycle arrest and promote cell apoptosis by regulating PI3 K/AKT signaling pathway.
引文
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[3]SI L,YANG X,YAN X,et al.Isoliquiritigenin induces apoptosis of human bladder cancer T24 cells via a cyclin-dependent kinase-independent mechanism[J].Oncol Lett,2017,14(1):241
[4]JI Y,CHEN S,LI K,et al.Signaling pathways in the development of infantile hemangioma[J].J Hematol Oncol,2014,7(1):13
[5]JI BC,ZHANG ZD,GUO WT,et al.Isoliquiritigenin blunts osteoarthritis by inhibition of bone resorption and angiogenesis in subchondral bone[J].Sci Rep,2018,8(1):1721
[6]HUANG YL,WEI F,ZHAO K,et al.Isoliquiritigenin inhibits colorectal cancer cells HCT-116 growth by suppressing the PI3K/AKT pathway[J].Open Life Sci,2017,12(1):300
[7]PENG F,TANG HL,LIU P,et al.Isoliquiritigenin modulates miR-374a/PTEN/Akt axis to suppress breast cancer tumorigenesis and metastasis[J].Sci Rep,2017,7(1):9022
[8]CHEN HY,HUANG TC,SHIEH TM,et al.Isoliquiritigenin induces autophagy and inhibits ovarian cancer cell growth[J].Int J Mol Sci,2017,18(10):2025
[9]王志强,张秀英,李文广,等.异甘草素抗肿瘤活性及初步机制研究[J].中国药理学通报,2015,31(8):1159
[10]MARTINI M,DE SANTIS MC,BRACCINI L,et al.PI3K/AKT signaling pathway and cancer:an updated review[J].Ann Med,2014,46(6):372
[11]LI Y,DAI YB,SUN JY,et al.Neuroglobin attenuates beta amyloid-induced apoptosis through inhibiting caspases activity by activating PI3K/Akt signaling pathway[J].J Mol Neurosci,2016,58(1):28
[12]LIN ML,CHEN SS,HUANG RY,et al.Suppression of PI3K/Akt signaling by synthetic bichalcone analog TSWU-CD4 induces ER stress-and Bax/Bak-mediated apoptosis of cancer cells[J].Apoptosis,2014,19(11):1637
[13]WU SY,WANG B,CHEN LF,et al.Clinical efficacy of propranolol in the treatment of hemangioma and changes in serum VEGF,b FGF and MMP-9[J].Exp Ther Med,2015,10(3):1079
[14]ZHAO HT,YUAN WL,WANG XK,et al.Expression and significance of apoptosis-related factors of mitochondrial pathway in different phases of infantile hemangioma[J].JHard Tissue Biol,2012,21(3):299
[15]陈志雄,张端莲,陕声国,等.Cyclin B1蛋白在血管瘤组织中的表达[J].中国组织化学与细胞化学杂志,2003,12(3):259
[16]吉毅,陈思源,李凯,等.PI3K/Akt信号通路的异常活化对婴幼儿血管瘤内皮细胞凋亡的抑制作用[J].中华小儿外科杂志,2014,35(2):93
[17]张晶,杨静,汤宏斌.异甘草素对人宫颈癌细胞增殖的抑制作用[J].中国药理学与毒理学杂志,2005,19(6):436
[18]刘方康,牛琼,王爱丽,等.异甘草素对胃癌SGC7901细胞侵袭能力的影响[J].天津医药,2015,43(11):1267