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Molecular Cancer
Published in: Molecular Cancer 1/2012

Open Access 01-12-2012 | Research

Inhibition of Phosphodiesterase-4 (PDE4) activity triggers luminal apoptosis and AKT dephosphorylation in a 3-D colonic-crypt model

Authors: Toshiyuki Tsunoda, Takeharu Ota, Takahiro Fujimoto, Keiko Doi, Yoko Tanaka, Yasuhiro Yoshida, Masahiro Ogawa, Hiroshi Matsuzaki, Masato Hamabashiri, Darren R Tyson, Masahide Kuroki, Shingo Miyamoto, Senji Shirasawa

Published in: Molecular Cancer | Issue 1/2012

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Abstract

Background

We previously established a three-dimensional (3-D) colonic crypt model using HKe3 cells which are human colorectal cancer (CRC) HCT116 cells with a disruption in oncogenic KRAS, and revealed the crucial roles of oncogenic KRAS both in inhibition of apoptosis and in disruption of cell polarity; however, the molecular mechanism of KRAS-induced these 3-D specific biological changes remains to be elucidated.

Results

Among the genes that were upregulated by oncogenic KRAS in this model, we focused on the phosphodiesterase 4B (PDE4B) of which expression levels were found to be higher in clinical tumor samples from CRC patients in comparison to those from healthy control in the public datasets of gene expression analysis. PDE4B2 was specifically overexpressed among other PDE4 isoforms, and re-expression of oncogenic KRAS in HKe3 cells resulted in PDE4B overexpression. Furthermore, the inhibition of PDE4 catalytic activity using rolipram reverted the disorganization of HCT116 cells into the normal physiologic state of the epithelial cell polarity by inducing the apical assembly of ZO-1 (a tight junction marker) and E-cadherin (an adherens junction marker) and by increasing the activity of caspase-3 (an apoptosis marker) in luminal cavities. Notably, rolipram reduced the AKT phosphorylation, which is known to be associated with the disruption of luminal cavity formation and CRC development. Similar results were also obtained using PDE4B2-shRNAs. In addition, increased expression of PDE4B mRNA was found to be correlated with relapsed CRC in a public datasets of gene expression analysis.

Conclusions

These results collectively suggested that PDE4B is upregulated by oncogenic KRAS, and also that the inhibition of PDE4 catalytic activity can induce both epithelial cell polarity and luminal apoptosis in CRC, thus highlighting the utility of our 3-D culture (3 DC) model for the KRAS-induced development of CRC in 3-D microenvironment. Indeed, using this model, we found that PDE4B is a promising candidate for a therapeutic target as well as prognostic molecular marker in CRC. Further elucidation of the signaling network of PDE4B2 in 3 DC would provide a better understanding of CRC in vivo.
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Literature
1.
O'Brien LE, Zegers MM, Mostov KE: Opinion: Building epithelial architecture: insights from three-dimensional culture models. Nat Rev Mol Cell Biol. 2002, 3: 531-537. 10.1038/nrm859CrossRefPubMed
2.
Griffith LG, Swartz MA: Capturing complex 3D tissue physiology in vitro. Nat Rev Mol Cell Biol. 2006, 7: 211-224.CrossRefPubMed
3.
Wodarz A, Nathke I: Cell polarity in development and cancer. Nat Cell Biol. 2007, 9: 1016-1024. 10.1038/ncb433CrossRefPubMed
4.
Shirasawa S, Furuse M, Yokoyama N, Sasazuki T: Altered growth of human colon cancer cell lines disrupted at activated Ki-ras. Science. 1993, 260: 85-88. 10.1126/science.8465203CrossRefPubMed
5.
Tsunoda T, Takashima Y, Fujimoto T, Koyanagi M, Yoshida Y, Doi K, Tanaka Y, Kuroki M, Sasazuki T, Shirasawa S: Three-dimensionally specific inhibition of DNA repair-related genes by activated KRAS in colon crypt model. Neoplasia. 2010, 12: 397-404.PubMedCentralCrossRefPubMed
6.
7.
Erdogan S, Houslay MD: Challenge of human Jurkat T-cells with the adenylate cyclase activator forskolin elicits major changes in cAMP phosphodiesterase (PDE) expression by up-regulating PDE3 and inducing PDE4D1 and PDE4D2 splice variants as well as down-regulating a novel PDE4A splice variant. Biochem J. 1997, 321 (Pt 1): 165-175.PubMedCentralCrossRefPubMed
8.
Conti M, Beavo J: Biochemistry and physiology of cyclic nucleotide phosphodiesterases: essential components in cyclic nucleotide signaling. Annu Rev Biochem. 2007, 76: 481-511. 10.1146/annurev.biochem.76.060305.150444CrossRefPubMed
9.
Houslay MD: Underpinning compartmentalised cAMP signalling through targeted cAMP breakdown. Trends Biochem Sci. 2009, 35: 91-100.CrossRefPubMed
10.
Houslay MD, Baillie GS, Maurice DH: cAMP-Specific phosphodiesterase-4 enzymes in the cardiovascular system: a molecular toolbox for generating compartmentalized cAMP signaling. Circ Res. 2007, 100: 950-966. 10.1161/01.RES.0000261934.56938.38CrossRefPubMed
11.
Li X, Baillie GS, Houslay MD: Mdm2 directs the ubiquitination of beta-arrestin-sequestered cAMP phosphodiesterase-4D5. J Biol Chem. 2009, 284: 16170-16182. 10.1074/jbc.M109.008078PubMedCentralCrossRefPubMed
12.
Bolger GB, Peden AH, Steele MR, MacKenzie C, McEwan DG, Wallace DA, Huston E, Baillie GS, Houslay MD: Attenuation of the activity of the cAMP-specific phosphodiesterase PDE4A5 by interaction with the immunophilin XAP2. J Biol Chem. 2003, 278: 33351-33363. 10.1074/jbc.M303269200CrossRefPubMed
13.
Millar JK, Pickard BS, Mackie S, James R, Christie S, Buchanan SR, Malloy MP, Chubb JE, Huston E, Baillie GS: DISC1 and PDE4B are interacting genetic factors in schizophrenia that regulate cAMP signaling. Science. 2005, 310: 1187-1191. 10.1126/science.1112915CrossRefPubMed
14.
Page CP, Spina D: Phosphodiesterase inhibitors in the treatment of inflammatory diseases. Handb Exp Pharmacol. 2011, 204: 391-414. 10.1007/978-3-642-17969-3_17CrossRefPubMed
15.
Marquette A, Andre J, Bagot M, Bensussan A, Dumaz N: ERK and PDE4 cooperate to induce RAF isoform switching in melanoma. Nat Struct Mol Biol. 2011, 18: 584-591. 10.1038/nsmb.2022CrossRefPubMed
16.
Keravis T, Lugnier C: Cyclic nucleotide phosphodiesterases (PDE) and peptide motifs. Curr Pharm Des. 2010, 16: 1114-1125. 10.2174/138161210790963760CrossRefPubMed
17.
Reich M, Liefeld T, Gould J, Lerner J, Tamayo P, Mesirov JP: GenePattern 2.0. Nat Genet. 2006, 38: 500-501. 10.1038/ng0506-500CrossRefPubMed
18.
Hong Y, Ho KS, Eu KW, Cheah PY: A susceptibility gene set for early onset colorectal cancer that integrates diverse signaling pathways: implication for tumorigenesis. Clin Cancer Res. 2007, 13: 1107-1114. 10.1158/1078-0432.CCR-06-1633CrossRefPubMed
19.
Mangoo-Karim R, Uchic M, Lechene C, Grantham JJ: Renal epithelial cyst formation and enlargement in vitro: dependence on cAMP. Proc Natl Acad Sci U S A. 1989, 86: 6007-6011. 10.1073/pnas.86.15.6007PubMedCentralCrossRefPubMed
20.
Miyoshi J, Takai Y: Molecular perspective on tight-junction assembly and epithelial polarity. Adv Drug Deliv Rev. 2005, 57: 815-855. 10.1016/j.addr.2005.01.008CrossRefPubMed
21.
Jin SL, Lan L, Zoudilova M, Conti M: Specific role of phosphodiesterase 4B in lipopolysaccharide-induced signaling in mouse macrophages. J Immunol. 2005, 175: 1523-1531.CrossRefPubMed
22.
Debnath J, Walker SJ, Brugge JS: Akt activation disrupts mammary acinar architecture and enhances proliferation in an mTOR-dependent manner. J Cell Biol. 2003, 163: 315-326. 10.1083/jcb.200304159PubMedCentralCrossRefPubMed
23.
Smith PG, Wang F, Wilkinson KN, Savage KJ, Klein U, Neuberg DS, Bollag G, Shipp MA, Aguiar RC: The phosphodiesterase PDE4B limits cAMP-associated PI3K/AKT-dependent apoptosis in diffuse large B-cell lymphoma. Blood. 2005, 105: 308-316. 10.1182/blood-2004-01-0240CrossRefPubMed
24.
Kenny HA, Dogan S, Zillhardt M, Zillhardt M, Metra AK, Yamada SD, Krausz T, Lengyel E: Organotypic models of metastasis: A three-dimensional culture mimicking the human peritoneum and omentum for the study of the early steps of ovarian cancer metastasis. Cancer Treat Res. 2009, 149: 335-351. 10.1007/978-0-387-98094-2_16PubMedCentralCrossRefPubMed
25.
Tsunoda T, Furusato B, Takashima Y, Ravulapalli S, Dobi A, Srivastava S, McLeod DG, Sesterhenn IA, Ornstein DK, Shirasawa S: The increased expression of periostin during early stages of prostate cancer and advanced stages of cancer stroma. Prostate. 2009, 69: 1398-1403. 10.1002/pros.20988CrossRefPubMed
26.
Fournier MV, Martin KJ, Kenny PA, Xhaja K, Bosch I, Yaswen P, Bissell MJ: Gene expression signature in organized and growth-arrested mammary acini predicts good outcome in breast cancer. Cancer Res. 2006, 66: 7095-7102. 10.1158/0008-5472.CAN-06-0515PubMedCentralCrossRefPubMed
27.
Martin KJ, Patrick DR, Bissell MJ, Fournier MV: Prognostic breast cancer signature identified from 3D culture model accurately predicts clinical outcome across independent datasets. PLoS One. 2008, 3: e2994 10.1371/journal.pone.0002994PubMedCentralCrossRefPubMed
28.
Bandrès E, Malumbres R, Cubedo E, Honorato B, Zarate R, Labarga A, Gabisu U, Sola JJ, Garcia-Foncillas J: A gene signature of 8 genes could identify the risk of recurrence and progression in Dukes' B colon cancer patients. Oncol Rep. 2007, 17: 1089-1094.PubMed
29.
Jaffe AB, Kaji N, Durgan J, Hall A: Cdc42 controls spindle orientation to position the apical surface during epithelial morphogenesis. J Cell Biol. 2008, 183: 625-633. 10.1083/jcb.200807121PubMedCentralCrossRefPubMed
30.
Martin-Belmonte F, Gassama A, Datta A, Yu W, Rescher U, Gerke V, Mostov K: PTEN-mediated apical segregation of phosphoinositides controls epithelial morphogenesis through Cdc42. Cell. 2007, 128: 383-397. 10.1016/j.cell.2006.11.051PubMedCentralCrossRefPubMed
31.
Kim J, Takeuchi H, Lam ST, Turner RR, Wang HJ, Kuo C, Foshag L, Bilchik AJ, Hoon DS: Chemokine receptor CXCR4 expression in colorectal cancer patients increases the risk for recurrence and for poor survival. J Clin Oncol. 2005, 23: 2744-2753.CrossRefPubMed
32.
Zeelenberg IS, Ruuls-Van Stalle L, Roos E: The chemokine receptor CXCR4 is required for outgrowth of colon carcinoma micrometastases. Cancer Res. 2003, 63: 3833-3839.PubMed
33.
Balko JM, Potti A, Saunders C, Stromberg A, Haura EB, Black EP: Gene expression patterns that predict sensitivity to epidermal growth factor receptor tyrosine kinase inhibitors in lung cancer cell lines and human lung tumors. BMC Genomics. 2006, 7: 289- 10.1186/1471-2164-7-289PubMedCentralCrossRefPubMed
34.
Park SJ, Ahmad F, Philp A, Baar K, Williams T, Luo H, Ke H, Rehmann H, Taussig R, Brown AL: Resveratrol ameliorates aging-related metabolic phenotypes by inhibiting cAMP phosphodiesterases. Cell. 2012, 148: 421-433. 10.1016/j.cell.2012.01.017PubMedCentralCrossRefPubMed
35.
Kim SW, Rai D, McKeller MR, Aguiar RC: Rational combined targeting of phosphodiesterase 4B and SYK in DLBCL. Blood. 2009, 113: 6153-6160. 10.1182/blood-2009-02-206128PubMedCentralCrossRefPubMed
36.
Meyers JA, Su DW, Lerner A: Chronic lymphocytic leukemia and B and T cells differ in their response to cyclic nucleotide phosphodiesterase inhibitors. J Immunol. 2009, 182: 5400-5411. 10.4049/jimmunol.0804255PubMedCentralCrossRefPubMed
37.
Sengupta R, Sun T, Warrington NM, Rubin JB: Treating brain tumors with PDE4 inhibitors. Trends Pharmacol Sci. 2011, 32: 337-344. 10.1016/j.tips.2011.02.015PubMedCentralCrossRefPubMed
38.
Chen TC, Wadsten P, Su S, Rawlinson N, Hofman FM, Hill CK, Schonthal AH: The type IV phosphodiesterase inhibitor rolipram induces expression of the cell cycle inhibitors p21(Cip1) and p27(Kip1), resulting in growth inhibition, increased differentiation, and subsequent apoptosis of malignant A-172 glioma cells. Cancer Biol Ther. 2002, 1: 268-276.CrossRefPubMed
39.
Murata K, Sudo T, Kameyama M, Fukuoka H, Muka M, Doki Y, Sasaki Y, Ishikawa O, Kimura Y, Imaoka S: Cyclic AMP specific phosphodiesterase activity and colon cancer cell motility. Clin Exp Metastasis. 2000, 18: 599-604. 10.1023/A:1011926116777CrossRefPubMed
40.
McEwan DG, Brunton VG, Baillie GS, Leslie NR, Houslay MD, Frame MC: Chemoresistant KM12C colon cancer cells are addicted to low cyclic AMP levels in a phosphodiesterase 4-regulated compartment via effects on phosphoinositide 3-kinase. Cancer Res. 2007, 67: 5248-5257. 10.1158/0008-5472.CAN-07-0097CrossRefPubMed
41.
Baba I, Shirasawa S, Iwamoto R, Okumura K, Tsunoda T, Nishioka M, Fukuyama K, Yamamoto K, Mekada E, Sasazuki T: Involvement of deregulated epiregulin expression in tumorigenesis in vivo through activated Ki-Ras signaling pathway in human colon cancer cells. Cancer Res. 2000, 60: 6886-6889.PubMed
42.
Tsunoda T, Takashima Y, Yoshida Y, Doi K, Tanaka Y, Fujimoto T, Machida T, Ota T, Koyanagi M, Kuroki M: Oncogenic KRAS Regulates miR-200c and miR-221/222 in a 3D-Specific Manner in Colorectal Cancer Cells. Anticancer Res. 2011, 31: 2453-2459.PubMed
43.
Koyanagi M, Nakabayashi K, Fujimoto T, Gu N, Baba I, Takashima Y, Doi K, Harada H, Kato N, Sasazuki T, Shirasawa S: ZFAT expression in B and T lymphocytes and identification of ZFAT-regulated genes. Genomics. 2008, 91: 451-457. 10.1016/j.ygeno.2008.01.009CrossRefPubMed
44.
Bolstad BM, Irizarry RA, Astrand M, Speed TP: A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics. 2003, 19: 185-193. 10.1093/bioinformatics/19.2.185CrossRefPubMed
45.
Golub TR, Slonim DK, Tamayo P, Huard C, Gaasenbeek M, Mesirov JP, Coller H, Loh ML, Downing JR, Caligiuri MA: Molecular classification of cancer: class discovery and class prediction by gene expression monitoring. Science. 1999, 286: 531-537. 10.1126/science.286.5439.531CrossRefPubMed
46.
Barber R, Baillie GS, Bergmann R, Shepherd MC, Sepper R, Houslay MD, Heeke GV: Differential expression of PDE4 cAMP phosphodiesterase isoforms in inflammatory cells of smokers with COPD, smokers without COPD, and nonsmokers. Am J Physiol Lung Cell Mol Physiol. 2004, 287: L332-L343. 10.1152/ajplung.00384.2003CrossRefPubMed
47.
Debnath J, Muthuswamy SK, Brugge JS: Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membrane cultures. Methods. 2003, 30: 256-268. 10.1016/S1046-2023(03)00032-XCrossRefPubMed
48.
Yoshida Y, Tsunoda T, Doi K, Tanaka Y, Fujimoto T, Machida T, Ota T, Koyanagi M, Takashima Y, Sasazuki T: KRAS-mediated up-regulation of RRM2 expression is essential for the proliferation of colorectal cancer cell lines. Anticancer Res. 2011, 31: 2535-2539.PubMed
Metadata
Title
Inhibition of Phosphodiesterase-4 (PDE4) activity triggers luminal apoptosis and AKT dephosphorylation in a 3-D colonic-crypt model
Authors
Toshiyuki Tsunoda
Takeharu Ota
Takahiro Fujimoto
Keiko Doi
Yoko Tanaka
Yasuhiro Yoshida
Masahiro Ogawa
Hiroshi Matsuzaki
Masato Hamabashiri
Darren R Tyson
Masahide Kuroki
Shingo Miyamoto
Senji Shirasawa
Publication date
01-12-2012
Publisher
BioMed Central
Published in
Molecular Cancer / Issue 1/2012
Electronic ISSN: 1476-4598
DOI
https://doi.org/10.1186/1476-4598-11-46

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