Abstract
Loss of actin stress fibers has been associated with cell transformation and metastasis. TGF-β induction of stress fibers in epithelial cells requires high molecular weight tropomyosins encoded by TPM1 and TPM2 genes. Here, we investigated the mechanism underlying the failure of TGF-β to induce stress fibers and inhibit cell migration in metastatic cells. RT–PCR analysis in carcinoma cell lines revealed a significant reduction in TPM1 transcripts in metastatic MDA-MB-231, MDA-MB-435 and SW620 cell lines. Treatment of these cells with demethylating agent 5-aza-2′-deoxycytidine (5-aza-dC) increased mRNA levels of TPM1 with no effect on TPM2. Importantly, 5-aza-dC treatment of MDA-MB-231 cells restored TGF-β induction of TPM1 and formation of stress fibers. Forced expression of TPM1 by using Tet-Off system increased stress fibers in MDA-MB-231 cells and reduced cell migration. A potential CpG island spanning the TPM1 proximal promoter, exon 1, and the beginning of intron 1 was identified. Bisulfite sequencing showed significant cytosine methylation in metastatic cell lines that correlated with a reduced expression of TPM1. Together these results suggest that epigenetic suppression of TPM1 may alter TGF-β tumor suppressor function and contribute to metastatic properties of tumor cells.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Accession codes
Abbreviations
- TGF-β:
-
transforming growth factor beta
- MAPK:
-
mitogen-activated protein kinase
- TPM:
-
tropomyosin
- 5-aza-dC:
-
5-aza-2′-deoxycytidine
References
Albo D, Berger DH, Wang TN, Hu X, Rothman V and Tuszynski GP . (1997). Surgery, 122, 493–499; discussion 499–500.
Antequera F and Bird A . (1993). PNAS, 90, 11995–11999.
Ayscough KR . (1998). Curr. Opin. Cell. Biol., 10, 102–111.
Bakin AV and Curran T . (1999). Science, 283, 387–390.
Bakin AV, Rinehart C, Tomlinson AK and Arteaga CL . (2002). J. Cell. Sci., 115, 3193–3206.
Bakin AV, Safina A, Rinehart C, Daroqui C, Darbary H and Helfman DM . (2004). Mol. Biol. Cell., 15, 4682–4694.
Bharadwaj S and Prasad GL . (2002). Cancer Lett., 183, 205–213.
Bhowmick NA, Ghiassi M, Bakin AV, Aakre M, Lundquist CA, Engel M, Arteaga CL and Moses HL . (2001). Mol. Biol. Cell, 12, 27–36.
Boyd J, Risinger JI, Wiseman RW, Merrick BA, Selkirk JK and Barrett JC . (1995). Proc. Natl. Acad. Sci. USA, 92, 11534–11538.
Braverman RH, Cooper HL, Lee HS and Prasad GL . (1996). Oncogene, 13, 537–545.
Chung KY, Agarwal A, Uitto J and Mauviel A . (1996). J. Biol. Chem., 271, 3272–3278.
Cross SH and Bird AP . (1995). Curr. Opin. Genet. Dev., 5, 309–314.
Derynck R, Akhurst RJ and Balmain A . (2001). Nat. Genet., 29, 117–129.
Dumont N, Bakin AV and Arteaga CL . (2003). J. Biol. Chem., 278, 3275–3285.
Edlund S, Landstrom M, Heldin CH and Aspenstrom P . (2002). Mol. Biol. Cell., 13, 902–914.
Feng XH, Lin X and Derynck R . (2000). EMBO J., 19, 5178–5193.
Franzen B, Linder S, Uryu K, Alaiya AA, Hirano T, Kato H and Auer G . (1996). Br. J. Cancer, 73, 909–913.
Frommer M, McDonald L, Millar D, Collis C, Watt F, Grigg G, Molloy P and Paul C . (1992). PNAS, 89, 1827–1831.
Gagos S, Hopwood VL, Iliopoulos D, Kostakis A, Karayannakos P, Yatzides H, Skalkeas GD and Pathak S . (1995). Anticancer Res., 15, 369–378.
Gimona M, Kazzaz JA and Helfman DM . (1996). Proc. Natl. Acad. Sci. USA, 93, 9618–9623.
Gimona M, Watakabe A and Helfman D . (1995). PNAS, 92, 9776–9780.
Greenwel P, Inagaki Y, Hu W, Walsh M and Ramirez F . (1997). J. Biol. Chem., 272, 19738–19745.
Hannigan M, Zhan L, Ai Y and Huang CK . (1998). Biochem. Biophys. Res. Commun., 246, 55–58.
Hendricks M and Weintraub H . (1981). Proc. Natl. Acad. Sci. USA, 78, 5633–5637.
Hendricks M and Weintraub H . (1984). Mol. Cell. Biol., 4, 1823–1833.
Hughes JA, Cooke-Yarborough CM, Chadwick NC, Schevzov G, Arbuckle SM, Gunning P and Weinberger RP . (2003). Glia, 42, 25–35.
Jaffe AB and Hall A . (2002). Adv. Cancer Res., 84, 57–80.
Janssen RA and Mier JW . (1997). Mol. Biol. Cell, 8, 897–908.
Leonardi CL, Warren RH and Rubin RW . (1982). Biochim. Biophys. Acta, 720, 154–162.
Li X, Cowell JK and Sossey-Alaoui K . (2004). Oncogene, 23, 1474–1480.
Masuda A, Takenaga K, Kondoh F, Fukami H, Utsumi K and Okayama H . (1996). Oncogene, 12, 2081–2088.
Miller SA, Dykes DD and Polesky HF . (1988). Nucleic Acids Res., 16, 1215.
Moustakas A and Kardassis D . (1998). Proc. Natl. Acad. Sci. USA, 95, 6733–6738.
Park IK, Lyu MA, Yeo SJ, Han TH and Kook YH . (2000). Biochim. Biophys. Acta, 1490, 302–310.
Pawlak G and Helfman DM . (2001). Curr. Opin. Genet. Dev., 11, 41–47.
Pawlak G and Helfman DM . (2002a). J. Biol. Chem., 277, 26927–26933, Epub 2002. May 14.
Pawlak G and Helfman DM . (2002b). Mol. Biol. Cell, 13, 336–347.
Pawlak G, McGarvey TW, Nguyen TB, Tomaszewski JE, Puthiyaveettil R, Malkowicz SB and Helfman DM . (2004). Int. J. Cancer, 110, 368–373.
Piek E, Moustakas A, Kurisaki A, Heldin CH and ten Dijke P . (1999). J. Cell. Sci., 112, 4557–4568.
Pittenger MF, Kazzaz JA and Helfman DM . (1994). Curr. Opin. Cell. Biol., 6, 96–104.
Raval GN, Bharadwaj S, Levine EA, Willingham MC, Geary RL, Kute T and Prasad GL . (2003). Oncogene, 22, 6194–6203.
Robbins J . (1998). Circ. Res., 82, 134–136.
Roberts AB and Wakefield LM . (2003). PNAS, 100, 8621–8623.
Sahai E, Olson MF and Marshall CJ . (2001). EMBO J., 20, 755–766.
Saito H, Tsujitani S, Oka S, Kondo A, Ikeguchi M, Maeta M and Kaibara N . (2000). Anticancer Res., 20, 4489–4493.
Shields JM, Mehta H, Pruitt K and Der CJ . (2002). Mol. Cell. Biol., 22, 2304–2317.
Temm-Grove CJ, Jockusch BM, Weinberger RP, Schevzov G and Helfman DM . (1998). Cell Motil. Cytoskeleton, 40, 393–407.
Varga AE, Leonardos L, Jackson P, Marreiros A and Cowled PA . (2004). Neoplasia, 6, 128–135.
Vial E, Sahai E and Marshall CJ . (2003). Cancer Cell., 4, 67–79.
Wakefield LM and Roberts AB . (2002). Curr. Opin. Genet. Dev., 12, 22–29.
Wang FL, Wang Y, Wong WK, Liu Y, Addivinola FJ, Liang P, Chen LB, Kantoff PW and Pardee AB . (1996). Cancer Res., 56, 3634–3637.
Yu J, Zhang L, Hwang PM, Rago C, Kinzler KW and Vogelstein B . (1999). Proc. Natl. Acad. Sci. USA, 96, 14517–14522.
Zhu WG, Srinivasan K, Dai Z, Duan W, Druhan LJ, Ding H, Yee L, Villalona-Calero MA, Plass C and Otterson GA . (2003). Mol. Cell. Biol., 23, 4056–4065.
Acknowledgements
We thank Bert Vogelstein for providing reagents, Erika VanDette for assistance with immunofluorescence studies; Dominic Smiraglia for aid in initial design of methylation primers; Andrew Leppard for assistance with graphics in Figure 5; Michael Higgins and John Cowell for stimulating discussions. This work was supported by PHS grant R01 CA95263 and USAMRMC grant DAMD17-02-01-0602 (to A.V.B.).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Varga, A., Stourman, N., Zheng, Q. et al. Silencing of the Tropomyosin-1 gene by DNA methylation alters tumor suppressor function of TGF-β. Oncogene 24, 5043–5052 (2005). https://doi.org/10.1038/sj.onc.1208688
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1208688
Keywords
This article is cited by
-
Tropomyosin1 isoforms underlie epithelial to mesenchymal plasticity, metastatic dissemination, and resistance to chemotherapy in high-grade serous ovarian cancer
Cell Death & Differentiation (2024)
-
Epigenetic silencing of TPM2 contributes to colorectal cancer progression upon RhoA activation
Tumor Biology (2016)
-
Hypoxia alters the recruitment of tropomyosins into the actin stress fibres of neuroblastoma cells
BMC Cancer (2015)
-
Expression of the actin-associated protein transgelin (SM22) is decreased in prostate cancer
Cell and Tissue Research (2010)
-
BMP-6 inhibits microRNA-21 expression in breast cancer through repressing δEF1 and AP-1
Cell Research (2009)