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BMC Cancer
Published in: BMC Cancer 1/2006

Open Access 01-12-2006 | Research article

Identification of genes regulated by Wnt/β-catenin pathway and involved in apoptosis via microarray analysis

Authors: Moli Huang, Yihua Wang, Daochun Sun, Hongxia Zhu, Yanbing Yin, Wei Zhang, Shangbin Yang, Lanping Quan, Jinfeng Bai, Shengqi Wang, Quan Chen, Songgang Li, Ningzhi Xu

Published in: BMC Cancer | Issue 1/2006

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Abstract

Background

Wnt/β-catenin pathway has critical roles in development and oncogenesis. Although significant progress has been made in understanding the downstream signaling cascade of this pathway, little is known regarding Wnt/β-catenin pathway modification of the cellular apoptosis.

Methods

To identify potential genes regulated by Wnt/β-catenin pathway and involved in apoptosis, we used a stably integrated, inducible RNA interference (RNAi) vector to specific inhibit the expression and the transcriptional activity of β-catenin in HeLa cells. Meanwhile, we designed an oligonucleotide microarray covering 1384 apoptosis-related genes. Using oligonucleotide microarrays, a series of differential expression of genes was identified and further confirmed by RT-PCR.

Results

Stably integrated inducible RNAi vector could effectively suppress β-catenin expression and the transcriptional activity of β-catenin/TCF. Meanwhile, depletion of β-catenin in this manner made the cells more sensitive to apoptosis. 130 genes involved in some important cell-apoptotic pathways, such as PTEN-PI3K-AKT pathway, NF-κB pathway and p53 pathway, showed significant alteration in their expression level after the knockdown of β-catenin.

Conclusion

Coupling RNAi knockdown with microarray and RT-PCR analyses proves to be a versatile strategy for identifying genes regulated by Wnt/β-catenin pathway and for a better understanding the role of this pathway in apoptosis. Some of the identified β-catenin/TCF directed or indirected target genes may represent excellent targets to limit tumor growth.
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Literature
1.
Huelsken J, Birchmeier W: New aspects of Wnt signaling pathways in higher vertebrates. Curr Opin Genet Dev. 2001, 11: 547-553. 10.1016/S0959-437X(00)00231-8.CrossRefPubMed
2.
Peifer M, Polakis P: Wnt signaling in oncogenesis and embryogenesis–a look outside the nucleus. Science. 2000, 287: 1606-1609. 10.1126/science.287.5458.1606.CrossRefPubMed
3.
Giles RH, van Es JH, Clevers H: Caught up in a Wnt storm: Wnt signaling in cancer. Biochim Biophys Acta. 2003, 1653: 1-24.PubMed
4.
Taipale J, Beachy PA: The Hedgehog and Wnt signalling pathways in cancer. Nature. 2001, 411: 349-354. 10.1038/35077219.CrossRefPubMed
5.
Moon RT, Kohn AD, De Ferrari GV, Kaykas A: WNT and beta-catenin signalling: diseases and therapies. Nat Rev Genet. 2004, 5: 691-701. 10.1038/nrg1427.CrossRefPubMed
6.
Li H, Pamukcu R, Thompson WJ: beta-Catenin signaling: therapeutic strategies in oncology. Cancer Biol Ther. 2002, 1: 621-625.CrossRefPubMed
7.
Chen S, Guttridge DC, You Z, Zhang Z, Fribley A, Mayo MW, Kitajewski J, Wang CY: Wnt-1 signaling inhibits apoptosis by activating beta-catenin/T cell factor-mediated transcription. J Cell Biol. 2001, 152: 87-96. 10.1083/jcb.152.1.87.CrossRefPubMedPubMedCentral
8.
9.
Hannon GJ, Rossi JJ: Unlocking the potential of the human genome with RNA interference. Nature. 2004, 431: 371-378. 10.1038/nature02870.CrossRefPubMed
10.
Mittal V: Improving the efficiency of RNA interference in mammals. Nat Rev Genet. 2004, 5: 355-365. 10.1038/nrg1323.CrossRefPubMed
11.
van de Wetering M, Oving I, Muncan V, Pon Fong MT, Brantjes H, van LD, Holstege FC, Brummelkamp TR, Agami R, Clevers H: Specific inhibition of gene expression using a stably integrated, inducible small-interfering-RNA vector. EMBO Rep. 2003, 4: 609-615. 10.1038/sj.embor.embor865.CrossRefPubMedPubMedCentral
12.
Okada H, Mak TW: Pathways of apoptotic and non-apoptotic death in tumour cells. Nat Rev Cancer. 2004, 4: 592-603. 10.1038/nrc1412.CrossRefPubMed
13.
Schmitt CA: Senescence, apoptosis and therapy–cutting the lifelines of cancer. Nat Rev Cancer. 2003, 3: 286-295. 10.1038/nrc1044.CrossRefPubMed
14.
Wang Y, Zhou X, Zhu H, Liu S, Zhou C, Zhang G, Xue L, Lu N, Quan L, Bai J, Zhan Q, Xu N: Overexpression of EB1 in human esophageal squamous cell carcinoma (ESCC) may promote cellular growth by activating beta-catenin/TCF pathway. Oncogene. 2005, 24: 6637-6645. 10.1038/sj.onc.1208819.CrossRefPubMed
15.
Lu JP, Ma ZC, Yang J, Huang J, Wang SR, Wang SQ: Ginsenoside Rg1-induced alterations in gene expression in TNF-alpha stimulated endothelial cells. Chin Med J (Engl). 2004, 117: 871-876.
16.
Ishitani T, Ninomiya-Tsuji J, Nagai S, Nishita M, Meneghini M, Barker N, Waterman M, Bowerman B, Clevers H, Shibuya H, Matsumoto K: The TAK1-NLK-MAPK-related pathway antagonizes signalling between beta-catenin and transcription factor TCF. Nature. 1999, 399: 798-802. 10.1038/21674.CrossRefPubMed
17.
Zhang X, Gaspard JP, Chung DC: Regulation of vascular endothelial growth factor by the Wnt and K-ras pathways in colonic neoplasia. Cancer Res. 2001, 61: 6050-6054.PubMed
18.
Howe LR, Subbaramaiah K, Chung WJ, Dannenberg AJ, Brown AM: Transcriptional activation of cyclooxygenase-2 in Wnt-1-transformed mouse mammary epithelial cells. Cancer Res. 1999, 59: 1572-577.PubMed
19.
Rockman SP, Currie SA, Ciavarella M, Vincan E, Dow C, Thomas RJ, Phillips WA: Id2 is a target of the beta-catenin/T cell factor pathway in colon carcinoma. J Biol Chem. 2001, 276: 45113-45119. 10.1074/jbc.M107742200.CrossRefPubMed
20.
Koh TJ, Bulitta CJ, Fleming JV, Dockray GJ, Varro A, Wang TC: Gastrin is a target of the beta-catenin/TCF-4 growth-signaling pathway in a model of intestinal polyposis. J Clin Invest. 2000, 106: 533-539.CrossRefPubMedPubMedCentral
21.
Brabletz T, Jung A, Dag S, Hlubek F, Kirchner T: beta-catenin regulates the expression of the matrix metalloproteinase-7 in human colorectal cancer. Am J Pathol. 1999, 155: 1033-1038.CrossRefPubMedPubMedCentral
22.
Crawford HC, Fingleton BM, Rudolph-Owen LA, Goss KJ, Rubinfeld B, Polakis P, Matrisian LM: The metalloproteinase matrilysin is a target of beta-catenin transactivation in intestinal tumors. Oncogene. 1999, 18: 2883-2891. 10.1038/sj.onc.1202627.CrossRefPubMed
23.
Wielenga VJ, Smits R, Korinek V, Smit L, Kielman M, Fodde R, Clevers H, Pals ST: Expression of CD44 in Apc and Tcf mutant mice implies regulation by the WNT pathway. Am J Pathol. 1999, 154: 515-523.CrossRefPubMedPubMedCentral
24.
Sun P, Xiong H, Kim TH, Ren B, Zhang Z: Positive Inter-Regulation between beta-catenin/T Cell Factor-4 Signaling and Endothelin-1 Signaling Potentiates Proliferation and Survival of Prostate Cancer Cells. Mol Pharmacol. 2006, 69: 520-531. 10.1124/mol.105.019620.CrossRefPubMed
25.
Ohigashi T, Mizuno R, Nakashima J, Marumo K, Murai M: Inhibition of Wnt signaling downregulates Akt activity and induces chemosensitivity in PTEN-mutated prostate cancer cells. Prostat. 2005, 62: 61-68. 10.1002/pros.20117.CrossRef
26.
Bodine PV, Billiard J, Moran RA, Ponce-de-Leon H, McLarney S, Mangine A, Scrimo MJ, Bhat RA, Stauffer B, Green J, Stein GS, Lian JB, Komm BS: The Wnt antagonist secreted frizzled-related protein-1 controls osteoblast and osteocyte apoptosis. J Cell Biochem. 2005, 96: 1212-1230. 10.1002/jcb.20599.CrossRefPubMed
27.
Mazieres J, You L, He B, Xu Z, Lee AY, Mikami I, McCormick F, Jablons DM: Inhibition of Wnt16 in human acute lymphoblastoid leukemia cells containing the t(1;19) translocation induces apoptosis. Oncogene. 2005, 24: 5396-5400. 10.1038/sj.onc.1208568.CrossRefPubMed
28.
Mikami I, You L, He B, Xu Z, Batra S, Lee AY, Mazieres J, Reguart N, Uematsu K, Koizumi K, Jablons DM: Efficacy of Wnt-1 monoclonal antibody in sarcoma cells. BMC Cancer. 2005, 5: 53-10.1186/1471-2407-5-53.CrossRefPubMedPubMedCentral
29.
He B, Reguart N, You L, Mazieres J, Xu Z, Lee AY, Mikami I, McCormick F, Jablons DM: Blockade of Wnt-1 signaling induces apoptosis in human colorectal cancer cells containing downstream mutations. Oncogene. 2005, 24: 3054-3058. 10.1038/sj.onc.1208511.CrossRefPubMed
30.
You L, He B, Xu Z, Uematsu K, Mazieres J, Fujii N, Mikami I, Reguart N, McIntosh JK, Kashani-Sabet M, McCormick F, Jablons DM: An anti-Wnt-2 monoclonal antibody induces apoptosis in malignant melanoma cells and inhibits tumor growth. Cancer Res. 2004, 64: 5385-5389. 10.1158/0008-5472.CAN-04-1227.CrossRefPubMed
31.
You L, He B, Xu Z, Uematsu K, Mazieres J, Mikami I, Reguart N, Moody TW, Kitajewski J, McCormick F, Jablons DM: Inhibition of Wnt-2-mediated signaling induces programmed cell death in non-small-cell lung cancer cells. Oncogene. 2004, 23: 6170-6174. 10.1038/sj.onc.1207844.CrossRefPubMed
32.
He B, You L, Uematsu K, Xu Z, Lee AY, Matsangou M, McCormick F, Jablons DM: A monoclonal antibody against Wnt-1 induces apoptosis in human cancer cells. Neoplasia. 2004, 6: 7-14.CrossRefPubMedPubMedCentral
33.
Veeramachaneni NK, Kubokura H, Lin L, Pippin JA, Patterson GA, Drebin JA, Battafarano RJ: Down-regulation of beta catenin inhibits the growth of esophageal carcinoma cells. J Thorac Cardiovasc Surg. 2004, 127: 92-98. 10.1016/j.jtcvs.2003.06.008.CrossRefPubMed
34.
Monga SP, Monga HK, Tan X, Mule K, Pediaditakis P, Michalopoulos GK: Beta-catenin antisense studies in embryonic liver cultures: role in proliferation, apoptosis, and lineage specification. Gastroenterology. 2003, 124: 202-216. 10.1053/gast.2003.50000.CrossRefPubMed
35.
Jiang Y, Zhou XD, Liu YK, Huang XW, Zhao Y, Xue Q, Sun RX, Chen J, Wu X: Antisense Tcf inhibits the neoplastic growth of liver cancer cells. J Cancer Res Clin Oncol. 2004, 130: 671-678. 10.1007/s00432-004-0580-9.CrossRefPubMed
36.
Ioannidis V, Beermann F, Clevers H, Held W: The beta-catenin–TCF-1 pathway ensures CD4(+)CD8(+) thymocyte survival. Nat Immunol. 2001, 2: 691-697. 10.1038/90623.CrossRefPubMed
37.
Almeida M, Han L, Bellido T, Manolagas SC, Kousteni S: Wnt Proteins Prevent Apoptosis of Both Uncommitted Osteoblast Progenitors and Differentiated Osteoblasts by {beta}-Catenin-dependent and -independent Signaling Cascades Involving Src/ERK and Phosphatidylinositol 3-Kinase/AKT. J Biol Chem. 2005, 280: 41342-41351. 10.1074/jbc.M502168200.CrossRefPubMed
38.
Yang F, Zeng Q, Yu G, Li S, Wang CY: Wnt/beta-catenin signaling inhibits death receptor-mediated apoptosis and promotes invasive growth of HNSCC. Cell Signal. 2006, 18: 679-687. 10.1016/j.cellsig.2005.06.015.CrossRefPubMed
39.
Hwang SG, Ryu JH, Kim IC, Jho EH, Jung HC, Kim K, Kim SJ, Chun JS: Wnt-7a causes loss of differentiated phenotype and inhibits apoptosis of articular chondrocytes via different mechanisms. J Biol Chem. 2004, 279: 26597-26604. 10.1074/jbc.M401401200.CrossRefPubMed
40.
Longo KA, Kennell JA, Ochocinska MJ, Ross SE, Wright WS, MacDougald OA: Wnt signaling protects 3T3-L1 preadipocytes from apoptosis through induction of insulin-like growth factors. J Biol Chem. 2002, 277: 38239-38244. 10.1074/jbc.M206402200.CrossRefPubMed
41.
Ueda Y, Hijikata M, Takagi S, Takada R, Takada S, Chiba T, Shimotohno K: Wnt/beta-catenin signaling suppresses apoptosis in low serum medium and induces morphologic change in rodent fibroblasts. Int J Cancer. 2002, 99: 681-688. 10.1002/ijc.10418.CrossRefPubMed
42.
de la Taille A, Rubin MA, Chen MW, Vacherot F, de Medina SG, Burchardt M, Buttyan R, Chopin D: Beta-catenin-related anomalies in apoptosis-resistant and hormone-refractory prostate cancer cells. Clin Cancer Res. 2003, 9: 1801-1807.PubMed
43.
Cobas M, Wilson A, Ernst B, Mancini SJ, MacDonald HR, Kemler R, Radtke F: Beta-catenin is dispensable for hematopoiesis and lymphopoiesis. J Exp Med. 2004, 199: 221-229. 10.1084/jem.20031615.CrossRefPubMedPubMedCentral
44.
Schulze A, Downward J: Navigating gene expression using microarrays–a technology review. Nat Cell Biol. 2001, 3: E190-195. 10.1038/35087138.CrossRefPubMed
45.
Dupont J, Renou JP, Shani M, Hennighausen L, LeRoith D: PTEN overexpression suppresses proliferation and differentiation and enhances apoptosis of the mouse mammary epithelium. J Clin Invest. 2002, 110: 815-825. 10.1172/JCI200213829.CrossRefPubMedPubMedCentral
46.
Vazquez F, Sellers WR: The PTEN tumor suppressor protein: an antagonist of phosphoinositide 3-kinase signaling. Biochim Biophys Acta. 2000, 1470: M21-35.PubMed
47.
Chang F, Lee JT, Navolanic PM, Steelman LS, Shelton JG, Blalock WL, Franklin RA, McCubrey JA: Involvement of PI3K/Akt pathway in cell cycle progression, apoptosis, and neoplastic transformation: a target for cancer chemotherapy. Leukemia. 2003, 17: 590-603. 10.1038/sj.leu.2402824.CrossRefPubMed
48.
Kucharczak J, Simmons MJ, Fan Y, Gelinas C: To be, or not to be NF-kappaB is the answer–role of Rel/NF-kappaB in the regulation of apoptosis. Oncogene. 2003, 22: 8961-8982. 10.1038/sj.onc.1207230.CrossRefPubMed
49.
Luo JL, Kamata H, Karin M: IKK/NF-kappaB signaling: balancing life and death–a new approach to cancer therapy. J Clin Invest. 2005, 115: 2625-2632. 10.1172/JCI26322.CrossRefPubMedPubMedCentral
50.
Vousden KH, Lu X: Live or let die: the cell's response to p53. Nat Rev Cancer. 2002, 2: 594-604. 10.1038/nrc864.CrossRefPubMed
51.
Bournat JC, Brown AM, Soler AP: Wnt-1 dependent activation of the survival factor NF-kappaB in PC12 cells. J Neurosci Res. 2000, 61: 21-32. 10.1002/1097-4547(20000701)61:1<21::AID-JNR3>3.0.CO;2-7.CrossRefPubMed
52.
Iwabuchi K, Bartel PL, Li B, Marraccino R, Fields S: Two cellular proteins that bind to wild-type but not mutant p53. Proc Natl Acad Sci USA. 1994, 91: 6098-6102. 10.1073/pnas.91.13.6098.CrossRefPubMedPubMedCentral
53.
Iwabuchi K, Li B, Massa HF, Trask BJ, Date T, Fields S: Stimulation of p53-mediated transcriptional activation by the p53-binding proteins, 53BP1 and 53BP2. J Biol Chem. 1998, 273: 26061-26068. 10.1074/jbc.273.40.26061.CrossRefPubMed
54.
Liang XQ, Cao EH, Zhang Y, Qin JF: A P53 target gene, PIG11, contributes to chemosensitivity of cells to arsenic trioxide. FEBS Lett. 2004, 569: 94-98. 10.1016/j.febslet.2004.05.057.CrossRefPubMed
55.
Chen Y, Sun R, Han W, Zhang Y, Song Q, Di C, Ma D: Nuclear translocation of PDCD5 (TFAR19): an early signal for apoptosis?. FEBS Lett. 2001, 509: 191-196. 10.1016/S0014-5793(01)03062-9.CrossRefPubMed
56.
Kissil JL, Cohen O, Raveh T, Kimchi A: Structure-function analysis of an evolutionary conserved protein, DAP3, which mediates TNF-alpha- and Fas-induced cell death. EMBO J. 1999, 18: 353-362. 10.1093/emboj/18.2.353.CrossRefPubMedPubMedCentral
57.
Tibbetts MD, Zheng L, Lenardo MJ: The death effector domain protein family: regulators of cellular homeostasis. Nat Immunol. 2003, 4: 404-409. 10.1038/ni0503-404.CrossRefPubMed
Metadata
Title
Identification of genes regulated by Wnt/β-catenin pathway and involved in apoptosis via microarray analysis
Authors
Moli Huang
Yihua Wang
Daochun Sun
Hongxia Zhu
Yanbing Yin
Wei Zhang
Shangbin Yang
Lanping Quan
Jinfeng Bai
Shengqi Wang
Quan Chen
Songgang Li
Ningzhi Xu
Publication date
01-12-2006
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2006
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-6-221

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