ZEB1上调神经细胞黏附分子促进胶质瘤细胞侵袭与转移
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摘要
胶质瘤是一种较为常见的颅内恶性肿瘤,其侵袭转移能力强,影响临床疗效。探讨胶质瘤发生侵袭转移的分子机制,寻找新的靶点干预胶质瘤侵袭转移是目前亟待解决的重大课题。我们前期研究中发现,神经细胞黏附分子(neuronal cell adhesion molecule,NRCAM)在各种胶质瘤细胞中的表达量均显著高于其在人正常星形胶质细胞(NHA)中的表达量(NRCAM在胶质瘤A172和T98G中的表达量分别是其在NHA中的2. 15和17. 63倍);且根据人类蛋白质组学数据库信息及qRT-PCR结果证实,NRCAM在胶质瘤组织中的表达量也显著高于其在正常组织中的表达。Kaplan-Meier分析提示,高表达的NRCAM与胶质瘤患者较差的预后正相关。在此基础上,通过生物信息学预测的方法结合双荧光素酶报告基因实验证实,转录因子锌指E盒结合蛋白1(ZEB1)能够增加NRCAM启动子活性,上调NRCAM mRNA和蛋白质水平的表达量。通过Transwell实验证实,在过表达ZEB1的胶质瘤细胞A172中,沉默NRCAM将抑制该细胞的侵袭能力。而在敲低ZEB1的胶质瘤细胞T98G中,过表达NRCAM将增加该细胞的侵袭能力。总之,NRCAM在胶质瘤中显著高表达且与患者较差的预后正相关。ZEB1转录上调NRCAM来增加胶质瘤细胞侵袭能力。
Glioma is a common type of intracranial malignant tumor. Its invasion and metastasis ability is strong,leading to poor prognosis. Understanding the molecular mechanism of invasion and metastasis of glioma could help finding poential targets for therapy. Previously,we found that neuronal cell adhesion molecule(NRCAM) is highly expressed in glioma cells,comparing with NHA cells. Online databank information and qRT-PCR results suggested that NRCAM expression was enhanced in glioma tissues,comparing with normal tissues. Furthermore,Kaplan-Meier analysis noted that NRCAM was positively correlated with poor outcome of glioma cancer patients. Bioinformatics analysis combining with dual-luciferase reporter assay showed that transcriptional factor zinc finger E-box binding homeobox 1(ZEB1)could enhance promoter activity of NRCAM and up-regulated its mRNA and protein expression.Moreover,transwell assays were performed to evaluate cell invasion ability. Results suggested that knock down of NRCAM inhibited invasion of ZEB1-overexpressed A172 glioma cells. Overexpression of NRCAM enhanced invasion of T98 G glioma cells in which ZEB1 was knocked down. In conclusion,NRCAM is highly expressed in glioma and positively correlated with poor outcome of glioma patients.ZEB1 transcriptionally up-regulated NRCAM to enhance the invasion of glioma cells.
Glioma is a common type of intracranial malignant tumor. Its invasion and metastasis ability is strong,leading to poor prognosis. Understanding the molecular mechanism of invasion and metastasis of glioma could help finding poential targets for therapy. Previously,we found that neuronal cell adhesion molecule(NRCAM) is highly expressed in glioma cells,comparing with NHA cells. Online databank information and qRT-PCR results suggested that NRCAM expression was enhanced in glioma tissues,comparing with normal tissues. Furthermore,Kaplan-Meier analysis noted that NRCAM was positively correlated with poor outcome of glioma cancer patients. Bioinformatics analysis combining with dual-luciferase reporter assay showed that transcriptional factor zinc finger E-box binding homeobox 1(ZEB1)could enhance promoter activity of NRCAM and up-regulated its mRNA and protein expression.Moreover,transwell assays were performed to evaluate cell invasion ability. Results suggested that knock down of NRCAM inhibited invasion of ZEB1-overexpressed A172 glioma cells. Overexpression of NRCAM enhanced invasion of T98 G glioma cells in which ZEB1 was knocked down. In conclusion,NRCAM is highly expressed in glioma and positively correlated with poor outcome of glioma patients.ZEB1 transcriptionally up-regulated NRCAM to enhance the invasion of glioma cells.
引文
[1]Siegel RL,Miller KD,Jemal A.Cancer statistics,2017[J].CACancer J Clin,2017,67(1):7-30
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[9]Yu JM,Sun W,Hua F,et al.BCL6 induces EMT by promoting the ZEB1-mediated transcription repression of E-cadherin in breast cancer cells[J].Cancer Lett,2015,365(2):190-200
[10]Yang X,Li L,Huang Q,et al.Wnt signaling through Snail1 and Zeb1 regulates bone metastasis in lung cancer[J].Am J Cancer Res,2015,5(2):748-755
[11]Wang Y,Wen M,Kwon Y,et al.CUL4A induces epithelialmesenchymal transition and promotes cancer metastasis by regulating ZEB1 expression[J].Cancer Res,2014,74(2):520-531
[12]Hortsch M.Structural and functional evolution of the L1 family:are four adhesion molecules better than one?[J].Mol Cell Neurosci,2000,15(1):1-10
[13]Górka B,Skubis-Zegado J,Mikula M,et al.Nr CAM,a neuronal system cell-adhesion molecule,is induced in papillary thyroid carcinomas[J].Br J Cancer,2007,97(4):531-538
[14]Conacci-Sorrell ME,Ben-Yedidia T,Shtutman M,et al.NrCAM is a target gene of the beta-catenin/LEF-1 pathway in melanoma and colon cancer and its expression enhances motility and confers tumorigenesis[J].Genes Dev,2002,16(16):2058-2072
[15]Conacci-Sorrell M,Kaplan A,Raveh S,et al.The shed ectodomain of Nr-CAM stimulates cell proliferation and motility,and confers cell transformation[J].Cancer Res,2005,65(24):11605-11612
[16]Conacci-Sorrell M,Zhurinsky J,Ben-Ze’ev A.The cadherincatenin adhesion system in signaling and cancer[J].J Clin Invest,2002,109(8):987-991
[17]Siebzehnrubl FA,Silver DJ,Tugertimur B,et al.The ZEB1pathway links glioblastoma initiation,invasion and chemoresistance[J].EMBO Mol Med,2013,5(8):1196-1212
[18]Sánchez-TillóE,Fanlo L,Siles L,et al.The EMT activator ZEB1 promotes tumor growth and determines differential response to chemotherapy in mantle cell lymphoma[J].Cell Death Differ,2014,21(2):247-257
[19]Galván JA,Zlobec I,Wartenberg M,et al.Expression of E-cadherin repressors SNAIL,ZEB1 and ZEB2 by tumour and stromal cells influences tumour-budding phenotype and suggests heterogeneity of stromal cells in pancreatic cancer[J].Br JCancer,2015,112(12):1944-1950
[20]Papadopoulou V,Postigo A,Sánchez-TillóE,et al.ZEB1 and CtBP form a repressive complex at a distal promoter element of the BCL6 locus[J].Biochem J,2010,427(3):541-550
[21]Takeyama Y,Sato M,Horio M,et al.Knockdown of ZEB1,a master epithelial-to-mesenchymal transition(EMT)gene,suppresses anchorage-independent cell growth of lung cancer cells[J].Cancer Lett,2010,296(2):216-224
[22]Zhang P,Sun Y,Ma L.ZEB1:at the crossroads of epithelialmesenchymal transition,metastasis and therapy resistance[J].Cell Cycle,2015,14(4):481-487
[23]Caramel J,Ligier M,Puisieux A.Pleiotropic roles for ZEB1 in cancer[J].Cancer Res,2018,78(1):30-35
[2]Chen W,Zheng R,Baade PD,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132
[3]Glass R,Synowitz M.CNS macrophages and peripheral myeloid cells in brain tumours[J].Acta Neuropathol,2014,128(3):347-362
[4]Omuro A,De Angelis LM.Glioblastoma and other malignant gliomas:a clinical review[J].JAMA,2013,310(17):1842-1850
[5]Rape A,Ananthanarayanan B,Kumar S.Engineering strategies to mimic the glioblastoma microenvironment[J].Adv Drug Deliv Rev,2014,79-80:172-183
[6]Iser IC,Pereira MB,Lenz G,et al.The epithelial-tomesenchymal transition-like process in glioblastoma:an updated systematic review and in silico investigation[J].Med Res Rev,2017,37(2):271-313
[7]Kahlert UD,Nikkhah G,Maciaczyk J.Epithelial-tomesenchymal(-like)transition as a relevant molecular event in malignant gliomas[J].Cancer Lett,2013,331(2):131-138
[8]Kalluri R,Weinberg RA.The basics of epithelial-mesenchymal transition[J].J Clin Invest,2009,119(6):1420-1428
[9]Yu JM,Sun W,Hua F,et al.BCL6 induces EMT by promoting the ZEB1-mediated transcription repression of E-cadherin in breast cancer cells[J].Cancer Lett,2015,365(2):190-200
[10]Yang X,Li L,Huang Q,et al.Wnt signaling through Snail1 and Zeb1 regulates bone metastasis in lung cancer[J].Am J Cancer Res,2015,5(2):748-755
[11]Wang Y,Wen M,Kwon Y,et al.CUL4A induces epithelialmesenchymal transition and promotes cancer metastasis by regulating ZEB1 expression[J].Cancer Res,2014,74(2):520-531
[12]Hortsch M.Structural and functional evolution of the L1 family:are four adhesion molecules better than one?[J].Mol Cell Neurosci,2000,15(1):1-10
[13]Górka B,Skubis-Zegado J,Mikula M,et al.Nr CAM,a neuronal system cell-adhesion molecule,is induced in papillary thyroid carcinomas[J].Br J Cancer,2007,97(4):531-538
[14]Conacci-Sorrell ME,Ben-Yedidia T,Shtutman M,et al.NrCAM is a target gene of the beta-catenin/LEF-1 pathway in melanoma and colon cancer and its expression enhances motility and confers tumorigenesis[J].Genes Dev,2002,16(16):2058-2072
[15]Conacci-Sorrell M,Kaplan A,Raveh S,et al.The shed ectodomain of Nr-CAM stimulates cell proliferation and motility,and confers cell transformation[J].Cancer Res,2005,65(24):11605-11612
[16]Conacci-Sorrell M,Zhurinsky J,Ben-Ze’ev A.The cadherincatenin adhesion system in signaling and cancer[J].J Clin Invest,2002,109(8):987-991
[17]Siebzehnrubl FA,Silver DJ,Tugertimur B,et al.The ZEB1pathway links glioblastoma initiation,invasion and chemoresistance[J].EMBO Mol Med,2013,5(8):1196-1212
[18]Sánchez-TillóE,Fanlo L,Siles L,et al.The EMT activator ZEB1 promotes tumor growth and determines differential response to chemotherapy in mantle cell lymphoma[J].Cell Death Differ,2014,21(2):247-257
[19]Galván JA,Zlobec I,Wartenberg M,et al.Expression of E-cadherin repressors SNAIL,ZEB1 and ZEB2 by tumour and stromal cells influences tumour-budding phenotype and suggests heterogeneity of stromal cells in pancreatic cancer[J].Br JCancer,2015,112(12):1944-1950
[20]Papadopoulou V,Postigo A,Sánchez-TillóE,et al.ZEB1 and CtBP form a repressive complex at a distal promoter element of the BCL6 locus[J].Biochem J,2010,427(3):541-550
[21]Takeyama Y,Sato M,Horio M,et al.Knockdown of ZEB1,a master epithelial-to-mesenchymal transition(EMT)gene,suppresses anchorage-independent cell growth of lung cancer cells[J].Cancer Lett,2010,296(2):216-224
[22]Zhang P,Sun Y,Ma L.ZEB1:at the crossroads of epithelialmesenchymal transition,metastasis and therapy resistance[J].Cell Cycle,2015,14(4):481-487
[23]Caramel J,Ligier M,Puisieux A.Pleiotropic roles for ZEB1 in cancer[J].Cancer Res,2018,78(1):30-35