TAp73调节肺腺癌细胞的血管生成能力依赖P53状态
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摘要
TAp73是P53家族的一员,能够调节肿瘤的生成、侵袭和转移。但是,TAp73调节肿瘤血管生成的作用备受争议。本研究将外源TAp73转染至P53基因表达状态不同的两株肺腺癌细胞系H1299(P53-null)和A549(wt P53)中,观察TAp73对肿瘤血管生成的作用并探讨与P53基因的关系。首先,使用RT-PCR和Western印迹验证转染效率。细胞划痕实验表明,TAp73在A549细胞中促进细胞迁移,而在H1299细胞中抑制细胞迁移。体外HUVEC血管形成结果表明,TAp73在A549细胞中促进细胞血管形成,而在H1299细胞中抑制细胞血管形成。同时,血管生成抑制蛋白1(VASH1)的表达水平,也分别升高或降低。本文研究结果表明,TAp73对肺腺癌细胞血管生成的作用依赖于P53基因的状态:在野生型P53基因存在时,TAp73促进血管生成,而在缺失P53基因的情况下,TAp73抑制血管生成。本研究对于TAp73作为肿瘤的潜在治疗靶点具有重要意义。
TAp73 is a member of the P53 family,which can regulate the formation,invasion and metastasis of tumor.However,the role of TAp73 in regulating tumor angiogenesis is controversial.In this study,we transfected TAp73 into lung adenocarcinoma cell lines H1299(P53-null) and A549(wt P53)with different P53 gene status to observe the effect of TAp73 on angiogenesis and to explore therelationship with P53 gene.For this purpose,we first verified the transfection efficiency using RT-PCR and Western blotting.Wound healing assay showed that TAp73 promoted cell migration in A549 cell line and inhibited cell migration in H1299 cell line after TAp73 was overexpressed.Similarly,in vitro HUVEC tube formation assay showed that TAp73 promoted the ability of tube formation in A549 cell line and inhibited the ability of tube formation in H1299 cell line after TAp73 was overexpressed.At the same time,the expression level of VASH1 also increased in A549 cells or decreased in H1299 cells,respectively.Our results suggest that the effect of TAp73 on angiogenesis in lung adenocarcinoma cells depends on the status of P53 gene.TAp73 promotes angiogenesis in the presence of wild-type P53 gene,and TAp73 inhibits angiogenesis in the absence of P53 gene,which is perhaps significant for TAp73 as a potential therapeutic target in tumors.
TAp73 is a member of the P53 family,which can regulate the formation,invasion and metastasis of tumor.However,the role of TAp73 in regulating tumor angiogenesis is controversial.In this study,we transfected TAp73 into lung adenocarcinoma cell lines H1299(P53-null) and A549(wt P53)with different P53 gene status to observe the effect of TAp73 on angiogenesis and to explore therelationship with P53 gene.For this purpose,we first verified the transfection efficiency using RT-PCR and Western blotting.Wound healing assay showed that TAp73 promoted cell migration in A549 cell line and inhibited cell migration in H1299 cell line after TAp73 was overexpressed.Similarly,in vitro HUVEC tube formation assay showed that TAp73 promoted the ability of tube formation in A549 cell line and inhibited the ability of tube formation in H1299 cell line after TAp73 was overexpressed.At the same time,the expression level of VASH1 also increased in A549 cells or decreased in H1299 cells,respectively.Our results suggest that the effect of TAp73 on angiogenesis in lung adenocarcinoma cells depends on the status of P53 gene.TAp73 promotes angiogenesis in the presence of wild-type P53 gene,and TAp73 inhibits angiogenesis in the absence of P53 gene,which is perhaps significant for TAp73 as a potential therapeutic target in tumors.
引文
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[14]Vikhanskaya F,Toh WH,Dulloo I,et al.p73 supports cellular growth through c-Jun-dependent AP-1 transactivation[J].Nat Cell Biol,2007,9(6):698-705
[15]Amelio I,Inoue S,Markert EK,et al.TAp73 opposes tumor angiogenesis by promoting hypoxia inducible factor 1αdegradation[J].Proc Natl Acad Sci USA,2015,112(1):226-231
[16]Stantic M,Sakil HA,Zirath H,et al.TAp73 suppresses tumor angiogenesis through repression of proangiogenic cytokines and HIF-1αactivity[J].Proc Natl Acad Sci USA,2015,112(1):220-225
[17]Dulloo I,Phang BH,Othman R,et al.Hypoxia-inducible TAp73 supports tumorigenesis by regulating the angiogenic transcriptome[J].Nat Cell Biol,2015,17(4):511-523
[18]Dulloo I,Hooi PB,Sabapathy K.Hypoxia-induced DNp73stabilization regulates Vegf-A expression and tumor angiogenesis similar to TAp73[J].Cell Cycle,2015,14(22):3533-3539
[19]Fernandez-Alonso R,Martin-Lopez M,Gonzalez-Cano L,et al.p73 is required for endothelial cell differentiation,migration and the formation of vascular networks regulating VEGF and TGFβsignaling[J].Cell Death Differ,2015,22(8):1287-1299
[20]Phang BH,Othman R,Bougeard G,et al.Amino-terminal p53mutations lead to expression of apoptosis proficient p47 and prognosticate better survival,but predispose to tumorigenesis[J].Proc Natl Acad Sci USA,2015,112(46):E6349-E6358
[21]Vikhanskaya F,Bani MR,Borsotti P,et al.p73 overexpression increases VEGF and reduces thrombospondin-1 production:implications for tumor angiogenesis[J].Oncogene,2001,20(50):7293-7300
[2]William WN Jr,Lin HY,Lee JJ,et al.Revisiting stage IIIB and IV non-small cell lung cancer:analysis of the surveillance,epidemiology,and end results data[J].Chest,2009,136(3):701-709
[3]Carmeliet P,Jain RK.Angiogenesis in cancer and other diseases[J].Nature,2000,407(6801):249-257
[4]Phang BH,Othman R,Bougeard G,et al.Amino-terminal p53mutations lead to expression of apoptosis proficient p47 and prognosticate better survival,but predispose to tumorigenesis[J].Proc Natl Acad Sci USA,2015,112(46):E6349-E6358
[5]Li AG,Piluso LG,Cai X,et al.An acetylation switch in p53mediates holo-TFIID recruitment[J].Mol Cell,2007,28(3):408-421
[6]Tanaka T,Ohkubo S,Tatsuno I,et al.h CAS/CSE1L associates with chromatin and regulates expression of select p53 target genes[J].Cell,2007,130(4):638-650
[7]Tomasini R,Tsuchihara K,Wilhelm M,et al.TAp73 knockout shows genomic instability with infertility and tumor suppressor functions[J].Genes Dev,2008,22(19):2677-2691
[8]Melino G,De Laurenzi V,Vousden KH.p73:Friend or foe in tumorigenesis[J].Nat Rev Cancer,2002,2(8):605-615
[9]Kaghad M,Bonnet H,Yang A,et al.Monoallelically expressed gene related to p53 at 1p36,a region frequently deleted in neuroblastoma and other human cancers.[J].Cell,1997,90(4):809-19.
[10]Kovalev S,Marchenko N,Swendeman S,et al.Expression level,allelic origin,and mutation analysis of the p73 gene in neuroblastoma tumors and cell lines[J].Cell Growth Differ,1998,9(11):897-903
[11]Du W,Jiang P,Mancuso A,et al.TAp73 enhances the pentose phosphate pathway and supports cell proliferation[J].Nat Cell Biol,2013,15(8):991-1000
[12]Donehower LA,Harvey M,Slagle BL,et al.Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumors[J].Nature,1992,356(6366):215-221
[13]Zaika AI,Kovalev S,Marchenko ND,et al.Overexpression of the wild type p73 gene in breast cancer tissues and cell lines[J].Cancer Res,1999,59(13):3257-3263
[14]Vikhanskaya F,Toh WH,Dulloo I,et al.p73 supports cellular growth through c-Jun-dependent AP-1 transactivation[J].Nat Cell Biol,2007,9(6):698-705
[15]Amelio I,Inoue S,Markert EK,et al.TAp73 opposes tumor angiogenesis by promoting hypoxia inducible factor 1αdegradation[J].Proc Natl Acad Sci USA,2015,112(1):226-231
[16]Stantic M,Sakil HA,Zirath H,et al.TAp73 suppresses tumor angiogenesis through repression of proangiogenic cytokines and HIF-1αactivity[J].Proc Natl Acad Sci USA,2015,112(1):220-225
[17]Dulloo I,Phang BH,Othman R,et al.Hypoxia-inducible TAp73 supports tumorigenesis by regulating the angiogenic transcriptome[J].Nat Cell Biol,2015,17(4):511-523
[18]Dulloo I,Hooi PB,Sabapathy K.Hypoxia-induced DNp73stabilization regulates Vegf-A expression and tumor angiogenesis similar to TAp73[J].Cell Cycle,2015,14(22):3533-3539
[19]Fernandez-Alonso R,Martin-Lopez M,Gonzalez-Cano L,et al.p73 is required for endothelial cell differentiation,migration and the formation of vascular networks regulating VEGF and TGFβsignaling[J].Cell Death Differ,2015,22(8):1287-1299
[20]Phang BH,Othman R,Bougeard G,et al.Amino-terminal p53mutations lead to expression of apoptosis proficient p47 and prognosticate better survival,but predispose to tumorigenesis[J].Proc Natl Acad Sci USA,2015,112(46):E6349-E6358
[21]Vikhanskaya F,Bani MR,Borsotti P,et al.p73 overexpression increases VEGF and reduces thrombospondin-1 production:implications for tumor angiogenesis[J].Oncogene,2001,20(50):7293-7300