Abstract
Overexpression of platelet-derived growth factor A-chain (PDGF-A) is clearly linked to autocrine and paracrine stimulation of malignant growth in many human cancers. We have shown previously that PDGF-A overexpression in choriocarcinoma, hepatoma and lung carcinoma cell lines is driven by the activity of a 66 bp enhancer element (ACE66) located approximately 7 kb upstream of the PDGF-A transcription start site. In this study, the ACE66 element is shown to be activated in JEG-3 choriocarcinoma cells through synergistic interactions between consensus DNA motifs for binding of vitamin D receptor, AP1 and ELK1. Binding of the vitamin D/retinoid-X receptor (VDR/RXRα) heterodimer to the ACE66 element was reconstituted in vitro with recombinant VDR/RXRα and with JEG-3 nuclear extract, and was verified in living JEG-3 cells by chromatin immunoprecipitation analysis. Transcriptional activity of the ACE66 element, as well as occupancy of the element by VDR/RXRα, was shown to be independent of stimulation with the hormonal VDR ligand, 1,25-dihydroxyvitamin D3. The jun kinase pathway of mitogen-activated protein kinase (MAPK) signaling was shown to activate the ACE66 enhancer, most likely through activation of factors binding to the AP1 element. These results identify a novel mechanism of transcriptional enhancement involving ligand-independent activity of the VDR/RXR heterodimer and MAPK signaling pathways that appears to play an important role in the overexpression of PDGF in many different settings of human malignancy.
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Abbreviations
- ACE:
-
A-chain cell-specific enhancer
- ChIP:
-
chromatin immunoprecipitation
- DMEM:
-
Dulbecco's modified Eagle's medium
- DR3:
-
direct repeat-3
- EIF:
-
ethylation interference footprinting
- EMSA:
-
electrophoretic mobility shift assay
- FBS:
-
fetal bovine serum
- MAPK:
-
mitogen-activated protein kinase
- MR:
-
mobility region
- PDGF:
-
platelet-derived growth factor
- RXR:
-
retinoid-X receptor
- VDR:
-
vitamin D receptor
- VDRE:
-
vitamin D response element
- 1,25-(OH)2D3, 1:
-
25-dihydroxyvitamin D3
References
Bhandari B, Wenzel UO, Marra F and Abboud HE . (1995). J. Biol. Chem., 270, 5541–5548.
Boyd KE and Farnham PJ . (1997). Mol. Cell. Biol., 17, 2529–2537.
Boyd KE and Farnham PJ . (1999). Mol. Cell. Biol., 19, 8393–8399.
Brown AJ, Dusso A and Slatopolsky E . (1999). Am. J. Physiol., 277, F157–F175.
Brown PH, Alani R, Preis LH, Szabo E and Birrer MJ . (1993). Oncogene, 8, 877–886.
Büning H, Baeurle PA and Zorbas H . (1995). Nucleic Acids Res., 23, 1443–1444.
Day FL, Rafty LA, Chesterman CN and Khachigian LM . (1999). J. Biol. Chem., 274, 23726–23733.
De Lean A, Munson PJ and Rodbard D . (1978). Am. J. Physiol., 235, E97–E102.
Dignam JD, Lebovitz RM and Roeder RG . (1983). Nucleic Acids Res., 11, 1475–1489.
Gashler AL, Bonthron DT, Madden SL, Rauscher III FJ, Collins T and Sukhatme VP . (1992). Proc. Natl. Acad. Sci. USA, 89, 10984–10988.
Gilbertson DG, Duff ME, West JW, Kelly JD, Sheppard PO, Hofstrand PD, Gao Z, Shoemaker K, Bukowski TR, Moore M, Feldhaus AL, Humes JM, Palmer TE and Hart CE . (2001). J. Biol. Chem., 276, 27406–27414.
Graham FL and Van der Eb AJ . (1973). Virology, 52, 456–467.
Guan Z, Buckman SY, Pentland AP, Templeton DJ and Morrison AR . (1998). J. Biol. Chem., 273, 12901–12908.
Hakim I and Bar-Shavit Z . (2003). J. Cell Biochem., 88, 986–998.
Hart LL and Davie JR . (2002). Biochem. Cell Biol., 80, 335–341.
Heldin C-H and Westermark B . (1999). Physiol. Rev., 79, 1283–1316.
Hennigan RF and Stambrook PJ . (2001). Mol. Biol. Cell, 12, 2352–2363.
Kaetzel DM . (2003). Cytokine Growth Factor Rev., 14, 427–446.
Kaetzel Jr DM, Maul RS, Liu B, Bonthron D, Fenstermaker RA and Coyne DW . (1994). Biochem. J., 301, 321–327.
Karin M . (1998). Ann. N.Y. Acad. Sci., 851, 139–146.
Kerner SA, Scott RA and Pike JW . (1989). Proc. Natl. Acad. Sci. USA, 86, 4455–4459.
Khachigian LM and Collins T . (1997). Circ. Res., 81, 457–461.
Khachigian LM, Santiago FS, Rafty LA, Chan OLW, Delbridge GJ, Bobik A and Collins T . (1999). Circ. Res., 84, 1258–1267.
Khachigian LM, Williams AJ and Collins T . (1995). J. Biol. Chem., 270, 27679–27686.
Koszewski NJ, Reinhardt TA and Horst RL . (1996). J. Steroid Biochem. Mol. Biol., 59, 377–388.
Koszewski NJ, Reinhardt TA, Langub MC, Malluche HH and Horst RL . (1998). Arch. Biochem. Biophys., 349, 388–396.
LaRochelle WJ, Jeffers M, McDonald WF, Chillakuru RA, Giese NA, Lokker NA, Sullivan C, Boldog FL, Yang M, Vernet C, Burgess CE, Fernandes E, Deegler LL, Rittman B, Shimkets J, Rothberg JM and Lichenstein HS . (2001). Nat. Cell Biol., 3, 517–521.
Lemon BD and Freedman LP . (1999). Curr. Opin. Genet. Dev., 9, 499–504.
Lin X, Wang Z, Gu L and Deuel TF . (1992). J. Biol. Chem., 267, 25614–25619.
Liu SM, Koszewski N, Lupez M, Malluche HH, Olivera A and Russell J . (1996). Mol. Endocrinol., 10, 206–215.
Ma D, Xing Z, Liu B, Pedigo N, Zimmer S, Bai Z, Postel E and Kaetzel DM . (2002). J. Biol. Chem., 277, 1560–1567.
Mangelsdorf DJ and Evans RM . (1995). Cell, 83, 841–850.
Maul RS, Zhang HX, Reid JD, Pedigo NG and Kaetzel DM . (1998). J. Biol. Chem., 273, 33239–33246.
Merika M and Thanos D . (2001). Curr. Opin. Genet. Dev., 11, 205–208.
Mulkins MA, Manolagas SC, Deftos LJ and Sussman HH . (1983). J. Biol. Chem., 258, 6219–6225.
Olive M, Krylov D, Echlin DR, Gardner K, Taparowsky E and Vinson C . (1997). J. Biol. Chem., 272, 18586–18594.
Pedigo N, Zhang H, Koszewski NJ and Kaetzel DM . (2003). Growth Factors, 21, 151–160.
Pietras K, Sjoblom T, Rubin K, Heldin CH and Ostman A . (2003). Cancer Cell, 3, 439–443.
Pike JW, Gooze LL and Haussler MR . (1980). Life Sci., 26, 407–414.
Rachez C, Lemon BD, Suldan Z, Bromleigh V, Gamble M, Näär AM, Erdjument-Bromage H, Tempst P and Freedman LP . (1999). Nature, 398, 824–832.
Rafty LA, Santiago FS and Khachigian LM . (2002). EMBO J., 21, 334–343.
Silverman ES, Khachigian LM, Lindner V, Williams AJ and Collins T . (1997). Am. J. Physiol., 42, H1415–H1426.
Silverman ES, Khachigian LM, Santiago FS, Williams AJ, Lindner V and Collins T . (1999). Am. J. Pathol., 155, 1311–1317.
Smeal T, Binetruy B, Mercola DA, Birrer M and Karin M . (1991). Nature, 354, 494–496.
Smith CL, Hager GL, Pike JW and Marx SJ . (1991). Mol. Endocrinol., 5, 867–878.
Sun T, Zhao Y, Mangelsdorf DJ and Simpson ER . (1998). Endocrinology, 139, 1684–1691.
Tanamura A, Nomura S, Kurauchi O, Furui T, Mizutani S and Tomoda Y . (1995). J. Obstet. Gynaecol., 21, 631–639.
Taplin ME and Balk SP . (2004). J. Cell Biochem., 91, 483–490.
Thanos D and Maniatis T . (1995). Cell, 83, 1091–1100.
Tolon RM, Castillo AI, Jimenez-Lara AM and Aranda A . (2000). Mol. Cell. Biol., 20, 8793–8802.
Umesono K, Murakami KK, Thompson CC and Evans RM . (1991). Cell, 65, 1255–1266.
Wang Z-Y, Madden SL, Deuel TF and Rauscher III FJ . (1992). J. Biol. Chem., 267, 21999–22002.
Xu L, Glass CK and Rosenfeld MG . (1999). Curr. Opin. Genet. Dev., 9, 140–147.
Acknowledgements
This work was supported by grants from the National Institutes of Health (HL62877, DK45518 and CA83237) and the Kentucky Lung Cancer Research Board. We thank David Bonthron and Tucker Collins for their kind support in the early phases of this project, particularly for providing many of the PDGF-A genomic clones and promoter vectors employed in this paper. We also thank Lise Binderup of Leo Pharmaceuticals for the gift of 1,25-(OH)2D3 used in these studies, Dennis Templeton for MEKK1 and c-JUN expression plasmids, Hollie Gravatte for technical assistance and Olivier Thibault for assistance with computer graphics.
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Pedigo, N., Zhang, H., Bruno, M. et al. A 5′-distal enhanceosome in the PDGF-A gene is activated in choriocarcinoma cells via ligand-independent binding of vitamin D receptor and constitutive jun kinase signaling. Oncogene 24, 2654–2666 (2005). https://doi.org/10.1038/sj.onc.1208336
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DOI: https://doi.org/10.1038/sj.onc.1208336