摘要
BACKGROUND Aberrant expression of stanniocalcin 2(STC2) is implicated in colon adenocarcinoma(COAD). A previous study identified that STC2 functions as a tumor promoter to drive development of some cancers, but the role of its overexpression in the development of COAD remains unclear.AIM To evaluate the regulation mechanism of STC2 overexpression in COAD.METHODS The expression of STC2 in COAD was assessed by TCGA COAD database and GEO(GSE50760). Methylation level of the STC2 promoter was evaluated with beta value in UALCAN platform, and the correlation between STC2 expression and survival rate was investigated with TCGA COAD. Transcription binding site prediction was conducted by TRANSFAC and LASAGNA, and a luciferase reporter system was used to identify STC2 promoter activity in several cell lines,including HEK293 T, NCM460, HT29, SW480, and HCT116. Western blotting was performed to evaluate the role of Sp1 on the expression of STC2.RESULTS The central finding of this work is that STC2 is overexpressed in COAD tissues and positively correlated with poor prognosis. Importantly, the binding site of the transcription factor Sp1 is widely located in the promoter region of STC2. A luciferase reporter system was successfully constructed to analyze the transcription activity of STC2, and knocking down the expression of Sp1 significantly inhibited the transcription activity of STC2. Furthermore, inhibition of Sp1 remarkably decreased protein levels of STC2.CONCLUSION Our data provide evidence that the transcription factor Sp1 is essential for the overexpression of STC2 in COAD through activation of promoter activity. Taken together, our finding provides new insights into the mechanism of oncogenic function of COAD by STC2.
BACKGROUND Aberrant expression of stanniocalcin 2(STC2) is implicated in colon adenocarcinoma(COAD). A previous study identified that STC2 functions as a tumor promoter to drive development of some cancers, but the role of its overexpression in the development of COAD remains unclear.AIM To evaluate the regulation mechanism of STC2 overexpression in COAD.METHODS The expression of STC2 in COAD was assessed by TCGA COAD database and GEO(GSE50760). Methylation level of the STC2 promoter was evaluated with beta value in UALCAN platform, and the correlation between STC2 expression and survival rate was investigated with TCGA COAD. Transcription binding site prediction was conducted by TRANSFAC and LASAGNA, and a luciferase reporter system was used to identify STC2 promoter activity in several cell lines,including HEK293 T, NCM460, HT29, SW480, and HCT116. Western blotting was performed to evaluate the role of Sp1 on the expression of STC2.RESULTS The central finding of this work is that STC2 is overexpressed in COAD tissues and positively correlated with poor prognosis. Importantly, the binding site of the transcription factor Sp1 is widely located in the promoter region of STC2. A luciferase reporter system was successfully constructed to analyze the transcription activity of STC2, and knocking down the expression of Sp1 significantly inhibited the transcription activity of STC2. Furthermore, inhibition of Sp1 remarkably decreased protein levels of STC2.CONCLUSION Our data provide evidence that the transcription factor Sp1 is essential for the overexpression of STC2 in COAD through activation of promoter activity. Taken together, our finding provides new insights into the mechanism of oncogenic function of COAD by STC2.
引文
1 Siegel RL,Miller KD,Jemal A.Cancer statistics,2018.CA Cancer J Clin 2018;68:7-30[PMID:29313949 DOI:10.3322/caac.21442]
2 Sobrero A,Grothey A,Iveson T,Labianca R,Yoshino T,Taieb J,Maughan T,Buyse M,AndréT,Meyerhardt J,Shields AF,Souglakos I,Douillard JY,Cervantes A.The hard road to data interpretation:3or 6 months of adjuvant chemotherapy for patients with stage III colon cancer?Ann Oncol 2018;29:1099-1107[PMID:29438451 DOI:10.1093/annonc/mdy064]
3 Vietti Violi N,Duran R,Demartines N,Sempoux C,Guiu B,Bize PE,Sala N,Halkic N,Knebel JF,Denys A.Local recurrence rate in patients with colorectal cancer liver metastasis after wedge resection or percutaneous radiofrequency ablation.Int J Hyperthermia 2018;34:1020-1028[PMID:29506424 DOI:10.1080/02656736.2017.1372644]
4 Suehiro Y,Wong CW,Chirieac LR,Kondo Y,Shen L,Webb CR,Chan YW,Chan AS,Chan TL,Wu TT,Rashid A,Hamanaka Y,Hinoda Y,Shannon RL,Wang X,Morris J,Issa JP,Yuen ST,Leung SY,Hamilton SR.Epigenetic-genetic interactions in the APC/WNT,RAS/RAF,and P53 pathways in colorectal carcinoma.Clin Cancer Res 2008;14:2560-2569[PMID:18451217 DOI:10.1158/1078-0432.CCR-07-1802]
5 Smith G,Carey FA,Beattie J,Wilkie MJ,Lightfoot TJ,Coxhead J,Garner RC,Steele RJ,Wolf CR.Mutations in APC,Kirsten-ras,and p53--alternative genetic pathways to colorectal cancer.Proc Natl Acad Sci U S A 2002;99:9433-9438[PMID:12093899 DOI:10.1073/pnas.122612899]
6 Colak S,Medema JP.Human colonic fibroblasts regulate stemness and chemotherapy resistance of colon cancer stem cells.Cell Cycle 2016;15:1531-1537[PMID:25483065 DOI:10.4161/15384101.2014.973321]
7 Buzzelli JN,Ouaret D,Brown G,Allen PD,Muschel RJ.Colorectal cancer liver metastases organoids retain characteristics of original tumor and acquire chemotherapy resistance.Stem Cell Res 2018;27:109-120[PMID:29414601 DOI:10.1016/j.scr.2018.01.016]
8 Luo CW,Pisarska MD,Hsueh AJ.Identification of a stanniocalcin paralog,stanniocalcin-2,in fish and the paracrine actions of stanniocalcin-2 in the mammalian ovary.Endocrinology 2005;146:469-476[PMID:15486227 DOI:10.1210/en.2004-1197]
9 Ishibashi K,Miyamoto K,Taketani Y,Morita K,Takeda E,Sasaki S,Imai M.Molecular cloning of a second human stanniocalcin homologue(STC2).Biochem Biophys Res Commun 1998;250:252-258[PMID:9753616 DOI:10.1006/bbrc.1998.9300]
10 Takei Y,Yamamoto H,Masuda M,Sato T,Taketani Y,Takeda E.Stanniocalcin 2 is positively and negatively controlled by 1,25(OH)(2)D(3)and PTH in renal proximal tubular cells.J Mol Endocrinol2009;42:261-268[PMID:19131501 DOI:10.1677/JME-08-0161]
11 Yahata K,Mori K,Mukoyama M,Sugawara A,Suganami T,Makino H,Nagae T,Fujinaga Y,Nabeshima Y,Nakao K.Regulation of stanniocalcin 1 and 2 expression in the kidney by klotho gene.Biochem Biophys Res Commun 2003;310:128-134[PMID:14511659 DOI:10.1016/j.bbrc.2003.08.131]
12 Marouli E,Graff M,Medina-Gomez C,Lo KS,Wood AR,Kjaer TR,Fine RS,Lu Y,Schurmann C,Highland HM,Rüeger S,Thorleifsson G,Justice AE,Lamparter D,Stirrups KE,Turcot V,Young KL,Winkler TW,Esko T,Karaderi T,Locke AE,Masca NG,Ng MC,Mudgal P,Rivas MA,Vedantam S,Mahajan A,Guo X,Abecasis G,Aben KK,Adair LS,Alam DS,Albrecht E,Allin KH,Allison M,Amouyel P,Appel EV,Arveiler D,Asselbergs FW,Auer PL,Balkau B,Banas B,Bang LE,Benn M,Bergmann S,Bielak LF,Blüher M,Boeing H,Boerwinkle E,B?ger CA,Bonnycastle LL,Bots ML,Bottinger EP,Bowden DW,Brandslund I,Breen G,Brilliant MH,Broer L,Burt AA,Butterworth AS,Carey DJ,Caulfield MJ,Chambers JC,Chasman DI,Chen YI,Chowdhury R,Christensen C,Chu AY,Cocca M,Collins FS,Cook JP,Corley J,Galbany JC,Cox AJ,Danesh J,Davies G,Deary IJ,Dedoussis G,Demerath EW,Dennis JG,Drenos F,Du M,Dunning AM,Easton DF,Ebeling T,Edwards TL,Ellinor PT,Elliott P,Evangelou E,Farmaki AE,Faul JD,Feitosa MF,Feng S,Ferrannini E,Ferrario MM,Ferrieres J,Florez JC,Ford I,Fornage M,Franks PW,Galesloot TE,Gan W,Gandin I,Gasparini P,Giedraitis V,Giri A,Girotto G,Gordon SD,Gorski M,Grarup N,Grove ML,Gudnason V,Gustafsson S,Hansen T,Harris KM,Harris TB,Hattersley AT,Hayward C,He L,Heid IM,Heikkil?K,Helgeland?,Hernesniemi J,Hewitt AW,Hocking LJ,Hollensted M,Holmen OL,Hovingh GK,Howson JM,Hoyng CB,Huang PL,Hveem K,Ikram MA,Ingelsson E,Jackson AU,Jansson JH,Jarvik GP,Jensen GB,Jhun MA,Jia Y,Jiang X,Johansson S,J?rgensen ME,J?rgensen T,Jousilahti P,Jukema JW,Kahali B,Kahn RS,K?h?nen M,Kamstrup PR,Kanoni S,Kaprio J,Karaleftheri M,Kardia SL,Karpe F,Kee F,Keeman R,Kiemeney LA,Kitajima H,Kluivers KB,Kocher T,Komulainen P,Kontto J,Kooner JS,Kooperberg C,Kovacs P,Kriebel J,Kuivaniemi H,Küry S,Kuusisto J,La Bianca M,Laakso M,Lakka TA,Lange EM,Lange LA,Langefeld CD,Langenberg C,Larson EB,Lee IT,Lehtim?ki T,Lewis CE,Li H,Li J,Lin H,Lin LA,Lin X,Lind L,Lindstr?m J,Linneberg A,Liu Y,Liu Y,Lophatananon A,Luan J,Lubitz SA,Lyytik?inen LP,Mackey DA,Madden PA,Manning AK,M?nnist?S,Marenne G,Marten J,Martin NG,Mazul AL,Meidtner K,Metspalu A,Mitchell P,Mohlke KL,Morgan A,Morris AD,Morris AP,Munroe PB,Nalls MA,Nauck M,Nelson CP,Neville M,Nielsen SF,Nikus K,Nj?lstad PR,Nordestgaard BG,Ntalla I,O'Connel JR,Oksa H,Loohuis LM,Ophoff RA,Owen KR,Packard CJ,Padmanabhan S,Palmer CN,Pasterkamp G,Patel AP,Pattie A,Pedersen O,Peissig PL,Peloso GM,Pennell CE,Perola M,Perry JA,Perry JR,Person TN,Pirie A,Polasek O,Posthuma D,Raitakari OT,Rasheed A,Rauramaa R,Reilly DF,Reiner AP,Renstr?m F,Ridker PM,Rioux JD,Robertson N,Robino A,Rolandsson O,Rudan I,Ruth KS,Saleheen D,Salomaa V,Samani NJ,Sandow K,Sapkota Y,Sattar N,Schmidt MK,Schreiner PJ,McCarthy MI,Willer CJ,Stefansson K,Borecki IB,Liu DJ,North KE,Pers TH,Lindgren CM,Oxvig C,Kutalik Z,Rivadeneira F,Loos RJ,Frayling TM,Hirschhorn JN,Deloukas P,Lettre G,Vozzi D,Walker M,Wang F,Wang CA,Wang S,Wang Y,Wareham NJ,Warren HR,Wessel J,Willems SM,Wilson JG,Witte DR,Woods MO,Wu Y,Yaghootkar H,Yao J,Yao P,Young R,Zeggini E,Zhan X,Zhang W,Zhao JH,Zhao W,Zhao W,Zheng H,Zhou W,Rotter JI,Boehnke M,Kathiresan S,Schulze MB,Scott RA,Shah S,Sim X,Sivapalaratnam S,Small KS,Smith AV,Smith JA,Southam L,Spector TD,Speliotes EK,Starr JM,Steinthorsdottir V,Stringham HM,Stumvoll M,Surendran P,Tansey KE,Tardif JC,Taylor KD,Teumer A,Thompson DJ,Thorsteinsdottir U,Thuesen BH,T?njes A,Tromp G,Trompet S,Tsafantakis E,Tuomilehto J,Tyrer JP,Uher R,Uitterlinden AG,Ulivi S,Varbo A,Varga TV,Varma R,Edwards DR,Vermeulen SH,Vestergaard H,Vitart V,Vogt TF,Bork-Jensen J,Frikke-Schmidt R,Gordon-Larsen P,Li-Gao R,Mook-Kanamori DO,Müller-Nurasyid M,'t Hart LM,Yerges-Armstrong LM,Heard-Costa NL,de Bakker PI,de Borst GJ,de Denus S,de Groot MC,de Mutsert R,den Hollander AI,Di Angelantonio E,van der Laan SW,Van Der Leij AR,van Duijn CM,van Schoor NM,van Setten J,'t Hart LM;EPIC-InterAct Consortium;CHD Exome+Consortium;ExomeBP Consortium;T2D-Genes Consortium;GoT2DGenes Consortium;Global Lipids Genetics Consortium;ReproGen Consortium;MAGIC Investigators.Rare and low-frequency coding variants alter human adult height.Nature 2017;542:186-190[PMID:28146470 DOI:10.1038/nature21039]
13 Jepsen MR,Kl?verpris S,Mikkelsen JH,Pedersen JH,Füchtbauer EM,Laursen LS,Oxvig C.Stanniocalcin-2 inhibits mammalian growth by proteolytic inhibition of the insulin-like growth factor axis.J Biol Chem 2015;290:3430-3439[PMID:25533459 DOI:10.1074/jbc.M114.611665]
14 Wu F,Li TY,Su SC,Yu JS,Zhang HL,Tan GQ,Liu JW,Wang BL.STC2 as a novel mediator for Mus81-dependent proliferation and survival in hepatocellular carcinoma.Cancer Lett 2017;388:177-186[PMID:27939696 DOI:10.1016/j.canlet.2016.11.039]
15 Na SS,Aldonza MB,Sung HJ,Kim YI,Son YS,Cho S,Cho JY.Stanniocalcin-2(STC2):A potential lung cancer biomarker promotes lung cancer metastasis and progression.Biochim Biophys Acta 2015;1854:668-676[PMID:25463045 DOI:10.1016/j.bbapap.2014.11.002]
16 Yang S,Ji Q,Chang B,Wang Y,Zhu Y,Li D,Huang C,Wang Y,Sun G,Zhang L,Guan Q,Xiang J,Wei W,Lu Z,Liao T,Meng J,Wang Z,Ma B,Zhou L,Wang Y,Yang G.STC2 promotes head and neck squamous cell carcinoma metastasis through modulating the PI3K/AKT/Snail signaling.Oncotarget 2017;8:5976-5991[PMID:27863406 DOI:10.18632/oncotarget.13355]
17 Jansen MP,Sas L,Sieuwerts AM,Van Cauwenberghe C,Ramirez-Ardila D,Look M,Ruigrok-Ritstier K,Finetti P,Bertucci F,Timmermans MM,van Deurzen CH,Martens JW,Simon I,Roepman P,Linn SC,van Dam P,Kok M,Lardon F,Vermeulen PB,Foekens JA,Dirix L,Berns EM,Van Laere S.Decreased expression of ABAT and STC2 hallmarks ER-positive inflammatory breast cancer and endocrine therapy resistance in advanced disease.Mol Oncol 2015;9:1218-1233[PMID:25771305 DOI:10.1016/j.molonc.2015.02.006]
18 Zhou H,Li YY,Zhang WQ,Lin D,Zhang WM,Dong WD.Expression of stanniocalcin-1 and stanniocalcin-2 in laryngeal squamous cell carcinoma and correlations with clinical and pathological parameters.PLoS One 2014;9:e95466[PMID:24743310 DOI:10.1371/journal.pone.0095466]
19 Esseghir S,Kennedy A,Seedhar P,Nerurkar A,Poulsom R,Reis-Filho JS,Isacke CM.Identification of NTN4,TRA1,and STC2 as prognostic markers in breast cancer in a screen for signal sequence encoding proteins.Clin Cancer Res 2007;13:3164-3173[PMID:17545519 DOI:10.1158/1078-0432.CCR-07-0224]
20 Wang YY,Li L,Zhao ZS,Wang HJ.Clinical utility of measuring expression levels of KAP1,TIMP1 and STC2 in peripheral blood of patients with gastric cancer.World J Surg Oncol 2013;11:81[PMID:23548070 DOI:10.1186/1477-7819-11-81]
21 Lin S,Guo Q,Wen J,Li C,Lin J,Cui X,Sang N,Pan J.Survival analyses correlate stanniocalcin 2overexpression to poor prognosis of nasopharyngeal carcinomas.J Exp Clin Cancer Res 2014;33:26[PMID:24606961 DOI:10.1186/1756-9966-33-26]
22 Wu J,Lai M,Shao C,Wang J,Wei JJ.STC2 overexpression mediated by HMGA2 is a biomarker for aggressiveness of high-grade serous ovarian cancer.Oncol Rep 2015;34:1494-1502[PMID:26165228DOI:10.3892/or.2015.4120]
23 Zhang ZH,Wu YG,Qin CK,Rong ZH,Su ZX,Xian GZ.Stanniocalcin 2 expression predicts poor prognosis of hepatocellular carcinoma.Oncol Lett 2014;8:2160-2164[PMID:25289096 DOI:10.3892/ol.2014.2520]
24 Wang H,Wu K,Sun Y,Li Y,Wu M,Qiao Q,Wei Y,Han ZG,Cai B.STC2 is upregulated in hepatocellular carcinoma and promotes cell proliferation and migration in vitro.BMB Rep 2012;45:629-634[PMID:23187001 DOI:10.5483/BMBRep.2012.45.11.086]
25 Law AY,Wong CK.Stanniocalcin-2 is a HIF-1 target gene that promotes cell proliferation in hypoxia.Exp Cell Res 2010;316:466-476[PMID:19786016 DOI:10.1016/j.yexcr.2009.09.018]
26 Chen B,Zeng X,He Y,Wang X,Liang Z,Liu J,Zhang P,Zhu H,Xu N,Liang S.STC2 promotes the epithelial-mesenchymal transition of colorectal cancer cells through AKT-ERK signaling pathways.Oncotarget 2016;7:71400-71416[PMID:27662663 DOI:10.18632/oncotarget.12147]
27 Law AY,Wong CK.Stanniocalcin-2 promotes epithelial-mesenchymal transition and invasiveness in hypoxic human ovarian cancer cells.Exp Cell Res 2010;316:3425-3434[PMID:20619259 DOI:10.1016/j.yexcr.2010.06.026]
28 Harper TA,Joshi AD,Elferink CJ.Identification of stanniocalcin 2 as a novel aryl hydrocarbon receptor target gene.J Pharmacol Exp Ther 2013;344:579-588[PMID:23269473 DOI:10.1124/jpet.112.201111]
29 Wang Y,Gao Y,Cheng H,Yang G,Tan W.Stanniocalcin 2 promotes cell proliferation and cisplatin resistance in cervical cancer.Biochem Biophys Res Commun 2015;466:362-368[PMID:26361149 DOI:10.1016/j.bbrc.2015.09.029]
30 Yuan Q,Zhan L,Zhang LL,Wang Q,Liu J,Jiang ZY,Hu XM,Yuan XC.Stanniocalcin 2 induces oxaliplatin resistance in colorectal cancer cells by upregulating P-glycoprotein.Can J Physiol Pharmacol2016;94:929-935[PMID:27245421 DOI:10.1139/cjpp-2015-0530]
31 Cheng H,Wu Z,Wu C,Wang X,Liow SS,Li Z,Wu YL.Overcoming STC2 mediated drug resistance through drug and gene co-delivery by PHB-PDMAEMA cationic polyester in liver cancer cells.Mater Sci Eng C Mater Biol Appl 2018;83:210-217[PMID:29208281 DOI:10.1016/j.msec.2017.08.075]
32 Cheng D,Zhao Y,Wang S,Jia W,Kang J,Zhu J.Human Telomerase Reverse Transcriptase(hTERT)Transcription Requires Sp1/Sp3 Binding to the Promoter and a Permissive Chromatin Environment.J Biol Chem 2015;290:30193-30203[PMID:26487723 DOI:10.1074/jbc.M115.662221]
33 Strand SH,Orntoft TF,Sorensen KD.Prognostic DNA methylation markers for prostate cancer.Int J Mol Sci 2014;15:16544-16576[PMID:25238417 DOI:10.3390/ijms150916544]
34 Kordowski F,Kolarova J,Schafmayer C,Buch S,Goldmann T,Marwitz S,Kugler C,Scheufele S,Gassling V,Németh CG,Brosch M,Hampe J,Lucius R,R?der C,Kalthoff H,Siebert R,Ammerpohl O,Reiss K.Aberrant DNA methylation of ADAMTS16 in colorectal and other epithelial cancers.BMCCancer 2018;18:796[PMID:30081852 DOI:10.1186/s12885-018-4701-2]
35 Law AY,Lai KP,Ip CK,Wong AS,Wagner GF,Wong CK.Epigenetic and HIF-1 regulation of stanniocalcin-2 expression in human cancer cells.Exp Cell Res 2008;314:1823-1830[PMID:18394600DOI:10.1016/j.yexcr.2008.03.001]
36 Wang Z,Wu X,Wang Y.A framework for analyzing DNA methylation data from Illumina Infinium HumanMethylation450 BeadChip.BMC Bioinformatics 2018;19:115[PMID:29671397 DOI:10.1186/s12859-018-2096-3]
37 Liu Z,Qi L,Li Y,Zhao X,Sun B.VEGFR2 regulates endothelial differentiation of colon cancer cells.BMC Cancer 2017;17:593[PMID:28854900 DOI:10.1186/s12885-017-3578-9]
38 Luo Y,Ye GY,Qin SL,Yu MH,Mu YF,Zhong M.ATAD2 Overexpression Identifies Colorectal Cancer Patients with Poor Prognosis and Drives Proliferation of Cancer Cells.Gastroenterol Res Pract 2015;2015:936564[PMID:26697062 DOI:10.1155/2015/936564]
39 Hou J,Wang Z,Xu H,Yang L,Yu X,Yang Z,Deng Y,Meng J,Feng Y,Guo X,Yang G.Stanniocalicin2 suppresses breast cancer cell migration and invasion via the PKC/claudin-1-mediated signaling.PLoSOne 2015;10:e0122179[PMID:25830567 DOI:10.1371/journal.pone.0122179]
40 Huang WY,Hsu SD,Huang HY,Sun YM,Chou CH,Weng SL,Huang HD.MethHC:a database of DNAmethylation and gene expression in human cancer.Nucleic Acids Res 2015;43:D856-D861[PMID:25398901 DOI:10.1093/nar/gku1151]
41 MacLeod MC,Powell KL,Kuzmin VA,Kolbanovskiy A,Geacintov NE.Interference of benzo[a]pyrene diol epoxide-deoxyguanosine adducts in a GC box with binding of the transcription factor Sp1.Mol Carcinog 1996;16:44-52[PMID:8634093 DOI:10.1002/(SICI)1098-2744(199605)16:1<44::AID-MC6>3.0.CO;2-O]
42 Jitrapakdee S,Slawik M,Medina-Gomez G,Campbell M,Wallace JC,Sethi JK,O'rahilly S,Vidal-Puig AJ.The peroxisome proliferator-activated receptor-gamma regulates murine pyruvate carboxylase gene expression in vivo and in vitro.J Biol Chem 2005;280:27466-27476[PMID:15917242 DOI:10.1074/jbc.M503836200]
