LPS刺激对人结肠癌SW480细胞基因表达的影响
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摘要
目的研究结直肠癌细胞炎症模型中基因表达的变化以及所涉及的信号通路。方法利用脂多糖(LPS)刺激SW480细胞1、3、6小时后提取RNA构建测序文库进行转录组测序,对表达有变化的基因归类,并进行京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)分析,对分析结果进行实时荧光定量PCR验证。结果发现LPS刺激SW480细胞1、3、6小时后,3个时间点表达均有显著变化的基因有250个。对这些差异表达的基因进行了KEGG信号通路富集分析发现利用脂多糖刺激构建的炎症模型里,TNF-α激活和NF-κB信号通路活化最显著。利用癌症基因组图谱(TCGA)数据库进一步对表达持续上调或下调的基因进行分析发现,部分基因在结肠癌标本中的表达趋势与LPS刺激结肠癌细胞引起的表达变化趋势一致。结论在利用脂多糖刺激构建的结肠癌细胞炎症模型中,炎症过程中的基因表达变化可能与结肠癌的发生发展密切相关。
Objective To investigate gene expression and involved signaling pathway of inflammation model of colorectal cell line. Methods Lipopolysaccharide(LPS) is used to stimulate sw480 cell line at 1, 3,and 6 hours. Total RNA from the sw480 cells was extracted after LPS exposure and then sequenced to construct library. The sequence data was classified and analyzed using KEGG database. Then, Quantitative Real-time PCR(q RT-PCR) was used to verify the analysis result. Results 250 m RNA transcripts with significant alteration at three time points were collected. The signaling pathways of TNF-α and NF-κB were obviously activated in the inflammation model of sw480 cell induced by LPS through KEGG Pathway enrichment analysis. Some of these significantly altered genes showed the same change tendency between our sequence data and The Cancer Genome Atlas database. Conclusions These changes of gene expression in the inflammation model induced by LPS might participate in the occurrence and development of colorectal cancer.
Objective To investigate gene expression and involved signaling pathway of inflammation model of colorectal cell line. Methods Lipopolysaccharide(LPS) is used to stimulate sw480 cell line at 1, 3,and 6 hours. Total RNA from the sw480 cells was extracted after LPS exposure and then sequenced to construct library. The sequence data was classified and analyzed using KEGG database. Then, Quantitative Real-time PCR(q RT-PCR) was used to verify the analysis result. Results 250 m RNA transcripts with significant alteration at three time points were collected. The signaling pathways of TNF-α and NF-κB were obviously activated in the inflammation model of sw480 cell induced by LPS through KEGG Pathway enrichment analysis. Some of these significantly altered genes showed the same change tendency between our sequence data and The Cancer Genome Atlas database. Conclusions These changes of gene expression in the inflammation model induced by LPS might participate in the occurrence and development of colorectal cancer.
引文
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[2]Van Emburgh B O,Sartore-Bianchi A,Di Nicolantonio F,et al.Acquired resistance to EGFR-targeted therapies in colorectal cancer[J].Mol Oncol.2014,8(6):1084-1094.
[3]Lawler M,Alsina D,Adams R,et al.Critical research gaps and recommendations to inform research prioritisation for more effective prevention and improved outcomes in colorectal cancer[J].Gut.2018,67(1):179-193.
[4]Wang T,Cai G,Qiu Y,et al.Structural segregation of gut microbiota between colorectal cancer patients and healthy volunteers[J].ISME J.2012,6(2):320-329
[5]Jobin C.Colorectal cancer:looking for answers in the microbiota[J].Cancer Discov.2013,3(4):384-387.
[6]Schwabe R F,Jobin C.The microbiome and cancer[J].Nat Rev Cancer.2013,13(11):800-812.
[7]Sears C L,Garrett W S.Microbes,microbiota,and colon cancer[J].Cell Host Microbe.2014,15(3):317-328
[8]Shen X J,Rawls J F,Randall T,et al.Molecular characterization of mucosal adherent bacteria and associations with colorectal adenomas[J].Gut Microbes.2010,1(3):138-147.
[9]Yu T,Guo F,Yu Y,et al.Fusobacterium nucleatum Promotes Chemoresistance to Colorectal Cancer by Modulating Autophagy[J].Cell.2017,170(3):548-563.
[10]Sender R,Fuchs S,Milo R,et al.Are We Really Vastly Outnumbered?Revisiting the Ratio of Bacterial to Host Cells in Humans[J].Cell.2016,164(3):337-340.
[11]Sun SC.The non-canonical NF-κB pathway in immunity and inflammation[J].Nat Rev Immunol.2017,17(9):545-558
[12]Wang L S,Kuo C T,Huang Y W,et al.Gene-Diet Interactions on Colorectal Cancer Risk[J].Curr Nutr Rep.2012,1(3):132-141.
[13]Herrinton L J,Liu L,Levin T R,et al.Incidence and mortality of colorectal adenocarcinoma in persons with inflammatory bowel disease from 1998 to 2010.Gastroenterology[J].2012,143(2):382-389.
[14]Kostic A D,Xavier R J,Gevers D.The microbiome in inflammatory bowel disease:current status and the future ahead[J].Gastroenterology.2014,146(6):1489-1499.
[15]Vijay-Kumar M,Aitken J D,Carvalho F A,et al.Metabolic syndrome and altered gut microbiota in mice lacking Toll-like receptor 5[J].Science.2010,328(5975):228-231.
[16]Garrett W S,Gallini C A,Yatsunenko T,et al.Enterobacteriaceae act in concert with the gut microbiota to induce spontaneous and maternally transmitted colitis[J].Cell Host Microbe.2010,8(3):292-300.
[17]Ekbom A,Helmick C,Zack M,et al.Ulcerative colitis and colorectal cancer.A population-based study[J].N Engl J Med.1990,323:1228-1233
[18]Soderlund S,Brandt L,Lapidus A,et al.Decreasing time-trends of colorectal cancer in a large cohort of patients with inflammatory bowel disease[J].Gastroenterology.2009,136(5):1561-1567
[19]Raetz C R,Whitfield C.Lipopolysaccharide endotoxins[J].Annu Rev Biochem.2002,71(71):635-700.
[20]Medzhitov R,Janeway C Jr.Innate immunity[J].N Engl J Med.2000,3;343(5):338-344.
[21]Beutler B,Cerami A.Tumor necrosis,cachexia,shock,and inflammation:a common mediator[J].Annu Rev Biochem.1988,57(57):505-518.
[22]Dinarello C A.Interleukin-1 and interleukin-1antagonism[J].Blood.1991,77(8):1627-52.
[23]任莉荣,舒钧.核因子κB的研究进展[J].医学综述.2011,17(6):814-816
[24]Shih V F,Tsui R,Caldwell A,et al.A single NFκBsystem for both canonical and non-canonical signaling[J].Cell Res,2011,21(1):86-102
[25]Parkhomchuk D,Borodina T,Amstislavskiy V,et al.Transcriptome analysis by strand-specific sequencing of complementary DNA[J].Nucleic Acids Res.2009,37(18):e123.
[26]Borst S E.The role of TNF-alpha in insulin resistance[J].Endocrine.2004,23(2-3):177-182.
[27]Moser B,Clark-Lewis I,Zwahlen R,et al.Neutrophilactivating properties of the melanoma growth-stimulatory activity[J].J Exp Med.1990,171(5):1797-1802.
[28]Cheng W L,Wang C S,Huang Y H,et al.Overexpression of Cxcl1 and its receptor CXCR2 promote tumor invasion in gastric cancer[J].Ann Oncol.2011,22(10):2267-2276.
[29]Singh S,Nannuru K C,Sadanandam A,et al.CXCR1 and CXCR2 enhances human melanoma tumourigenesis,growth and invasion[J].Br J Cancer.2009,100(10):1638-1646
[30]Kawanishi H,Matsui Y,Ito M,et al.Secreted Cxcl1 is a potential mediator and marker of the tumor invasion of bladder cancer[J].Clin Cancer Res.2008,14(9):2579-2587.
[31]Wang D,Wang H,Brown J,et al.Cxcl1 induced by prostaglandin E2 promotes angiogenesis in colorectal cancer[J].J Exp Med.2006,203(4):941-951.
[32]Ponta H,Sherman L,Herrlich P A.CD44:From adhesion molecules to signalling regulators[J].Nat Rev Mol Cell Biol.2003,4(1):33-45.
[33]Zhao P,Damerow M S,Stern P,et al.CD44 promotes Kras-dependent lung adenocarcinoma[J].Oncogene.2013,32(43):5186-5190.
[34]Okano M,Yamamoto H,Ohkuma H,et al.Significance of INHBA expression in human colorectal cancer[J].Oncol Rep.2013,30(6):2903-2908.
[35]Seder C W,Hartojo W,Lin L,et al.INHBA overexpression promotes cell proliferation and may be epigenetically regulated in esophageal adenocarcinoma[J].J Thorac Oncol.2009,4(4):455-462.
[36]Keeley E C,Mehrad B,Strieter R M.Chemokines as mediators of tumor angiogenesis and neovascularization[J].Experimental Cell Research.2011,317(5):685-690.
[37]Gálvez B G,Genís L,Matías-Román S,et al.Membrane type 1-matrix metalloproteinase is regulated by chemokines monocyte-chemoattractant protein-1/CCL2and interleukin-8/CXCL8 in endothelial cells during angiogenesis[J].The Journal of Biological Chemistry.2005,2 80(2):1292-1298.