藻蓝蛋白对非小细胞肺癌H1299生长、迁移和凋亡的功能影响
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摘要
藻蓝蛋白(C-phycocyanin,C-PC)是一种从顿顶螺旋藻中提取的天然水溶性无毒的活性蛋白。采用MTT法、克隆形成实验、流式细胞术、划痕实验等探究不同浓度的藻蓝蛋白对人类非小细胞肺癌H1299细胞活力、凋亡、迁移等的影响。结果表明:藻蓝蛋白能显著(p<0.05)降低H1299细胞存活率,通过下调周期蛋白cyclinA、CDK2,上调周期抑制因子P21使细胞周期阻滞在S期;藻蓝蛋白处理使细胞形态发生改变,同时通过调控bad、bax、bcl-xL、bcl-2等凋亡相关基因的表达促进细胞凋亡;体外划痕实验表明藻蓝蛋白通过调控基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)和基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)抑制细胞迁移。研究结果表明藻蓝蛋白能够通过细胞周期S期阻滞,调控细胞周期、凋亡和迁移相关基因的表达而抑制非小细胞肺癌H1299细胞的生长和迁移,进而诱导细胞凋亡,为进一步开发功能食品奠定坚实的理论基础。
Phycocyanin(C-PC)is a natural water-soluble and nontoxic functional protein extracted from Spirulina platensis. In this study,the effects of C-PC on proliferation,apoptosis and migration in non-small cell lung cancer H1299 cells by using MTT assay,cloning formation assay,flow cytometry and wound-healing assay. The results showed that phycocyanin could significantly(p<0.05)decrease the cell survival rate. C-PC could induce S phase arrest by regulating the expression levels of cell cycle related proteins cyclinA,CDK2,and P21.Furthermore,C-PC induced apoptosis of H1299 cells by regulating apoptotic genes including bad,bax,bcl-xL,and bcl-2.The results of wound-healing assay showed that C-PC inhibited cell migration through regulating MMP-9(matrix metalloproteinase-9)and MMP-2(matrix metalloproteinase-2). These results indicated that C-PC could induce cell cycle S phase arrest,regulate the expression of cell cycle,migration and apoptosis genes,resulting in the growth inhibition and apoptosis of H1299 cells,which would provide a theoretical basis for the development of novel natural functional substances and lung cancer treatment.
Phycocyanin(C-PC)is a natural water-soluble and nontoxic functional protein extracted from Spirulina platensis. In this study,the effects of C-PC on proliferation,apoptosis and migration in non-small cell lung cancer H1299 cells by using MTT assay,cloning formation assay,flow cytometry and wound-healing assay. The results showed that phycocyanin could significantly(p<0.05)decrease the cell survival rate. C-PC could induce S phase arrest by regulating the expression levels of cell cycle related proteins cyclinA,CDK2,and P21.Furthermore,C-PC induced apoptosis of H1299 cells by regulating apoptotic genes including bad,bax,bcl-xL,and bcl-2.The results of wound-healing assay showed that C-PC inhibited cell migration through regulating MMP-9(matrix metalloproteinase-9)and MMP-2(matrix metalloproteinase-2). These results indicated that C-PC could induce cell cycle S phase arrest,regulate the expression of cell cycle,migration and apoptosis genes,resulting in the growth inhibition and apoptosis of H1299 cells,which would provide a theoretical basis for the development of novel natural functional substances and lung cancer treatment.
引文
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[25]卢科莲,喻小兰,夏纪毅,等.黄芩素对宫颈癌Hela细胞增殖的抑制作用及机制探讨[J].山东医药,2014,54(27):4-7.
[2]李媛秋,么鸿雁.肺癌主要危险因素的研究进展[J].中国肿瘤,2016,25(10):782-786.
[3]Wan M X,Wang Z Y,Zhang Z,et al.A novel paradigm for the high-efficient production of phycocyanin from Galdieria sulphuraria[J].Bioresource Technology,2016,218:272-278.
[4]Dong C,Tang A,Zhao J,et al.ApcD is necessary for efficient energy transfer from phycobilisomes to photosystem I and helps to prevent photoinhibition in the cyanobacterium Synechococcus sp.PCC 7002[J].Biochimica Biophysica Acta,2009,1787(9):1122-1128.
[5]欧瑜,郑姗,林琳.藻蓝蛋白对体外诱导HepG2细胞损伤的保护作用[J].药物生物技术,2009,16(5):435-438.
[6]Liu Q,Huang Y,Zhang R,et al.Medical application of Spirulina platensis derived C-Phycocyanin[J].Evidence-Based Complementary and Alternative Medicine,2016,2016:7803846.
[7]张成,潘若望,李坤鹏,等.藻蓝蛋白抑制人结肠癌SW480细胞增殖的初步研究[J].中国微生态学杂志,2015,27(2):146-150.
[8]Ravi M,Tentu S,Baskar G,et al.Molecular mechanism of anti-cancer activity of phycocyanin in triple-negative breast cancer cells[J].BMC Cancer,2015,15(1):768.
[9]Liao G,Bing G,Gao Y,et al.Phycocyanin inhibits tumorigenic potential of pancreatic cancer cells:Role of apoptosis and autophagy[J].Scientific Reports,2016,6:34564.
[10]李冰,杨帆,吕丛仪,等.藻蓝蛋白协同全反式维甲酸影响人宫颈癌HeLa细胞的增殖和凋亡[J].中国肿瘤生物治疗杂志,2014,21(5):516-520.
[11]Pan R,Lu R,Zhang Y,et al.Spirulina,phycocyanin induces differential protein expression and apoptosis in SKOV-3 cells[J].International Journal of Biological Macromolecules,1994,81(3):746-749.
[12]Gao Y,Liao G,Xiang C,et al.Effects of phycocyanin on INS-1 pancreatic β-cell mediated by PI3K/Akt/FoxO1 signaling pathway[J].International Journal of Biological Macromolecules,2015,83:185-194.
[13]应俊,潘若望,王茂峰,等.藻蓝蛋白诱导喉癌HEP-2细胞凋亡的实验研究[J].中国病理生理杂志,2015,31(7):1189-1196.
[14]张成武,刘宇峰.螺旋藻藻蓝蛋白对人血癌细胞株HL-60,K-562和U-937的生长影响[J].海洋科学,2000,24(1):45-48.
[15]Basha O M,Hafez R A,El-Ayouty Y M,et al.C-phycocyanin inhibits cell proliferation and may induce apoptosis in human HepG2 cells[J].Egyptian Journal of Immunology,2008,15(2):161-167.
[16]Jiang L,Wang Y,Liu G,et al.C-phycocyanin exerts anti-cancer effects via the MAPK signaling pathway in MDA-MB-231 cells[J].Cancer Cell International,2018,18(1):12.
[17]Li B,Gao M H,Lv C Y,et al.Study of the synergistic effects of all-transretinoic acid and C-phycocyanin on the growth and apoptosis of A549 cells[J].European Journal of Cancer Prevention,2016,25(2):97-101.
[18]Bingula R,Dupuis C,Pichon C,et al.Study of the effects of betaine and/or C-phycocyanin on the growth of lung cancer A549 cells in vitro and in vivo[J].Journal of Oncology,2016,2016(1):8162952.
[19]郝帅,闫燕,李爽,等.食用色素藻蓝蛋白对A549非小细胞肺癌体外增殖和凋亡的影响[J].食品科学,2019,40(3):158-164.
[20]Fan W,Sun L,Zhou J Q,et al.Marsdenia tenacissima extract induces G0/G1 cell cycle arrest in human esophageal carcinoma cells by inhibiting mitogen-activated protein kinase(MAPK)signaling pathway[J].Chinese Journal of Natural Medicines,2015,13(6):428-437.
[21]Zang Q Q,Zhang L,Gao N,et al.Ophiopogonin D inhibits cell proliferation,causes cell cycle arrest at G2/M,and induces apoptosis in human breast carcinoma MCF-7 cells[J].Journal of Integrative Medicine,2016,14(1):51-59.
[22]薛洋,周清华,张尚福,等.MMP-2、MMP-9在肺癌中的表达及其与肺癌转移和预后关系的研究[J].华西医学,2008(2):225-227.
[23]?ukaszewicz-Zajac M,Mroczko B,Guzińska-Ustymowicz K,et al.Matrix metalloproteinase 2(MMP-2)and their tissue inhibitor 2(TIMP-2)in gastric cancer patients[J].Advances in Medical Sciences,2013,58(2):235-243.
[24]万双艳,万义增,郭莲怡.姜黄素对宫颈癌组织OPN及MMP-9表达的影响[J].中国妇幼保健,2014,29(33):5490-5492.
[25]卢科莲,喻小兰,夏纪毅,等.黄芩素对宫颈癌Hela细胞增殖的抑制作用及机制探讨[J].山东医药,2014,54(27):4-7.