Expression Profiles of Apoptosis Genes in Mammary Epithelial Cells

  • Seol, Myung Bok (Department of Applied Bioscience and Biotechnology, Biotechnology Research Institute, Institute of Agricultural Science and Technology, Chonnam National University) ;
  • Bong, Jin Jong (Department of Applied Bioscience and Biotechnology, Biotechnology Research Institute, Institute of Agricultural Science and Technology, Chonnam National University) ;
  • Baik, Myunggi (Department of Applied Bioscience and Biotechnology, Biotechnology Research Institute, Institute of Agricultural Science and Technology, Chonnam National University)
  • Received : 2005.03.03
  • Accepted : 2005.05.02
  • Published : 2005.08.31

Abstract

To investigate apoptosis in HC11 mammary epithelial cells, we compared the gene expression profiles of actively growing and serum-starved apoptotic cells using a mouse apoptosis gene array and $^{33}P$-labeled cDNA prepared from the RNA of the two cultures. Analysis of the arrays showed that expression of several genes such as clusterin, secreted frizzled related protein mRNA (sFRP-1), CREB-binding protein (CBP), and others was higher in the apoptotic cells whereas expression of certain genes including survivin, cell division cycle 2 homolog A (CDC2), and cyclin A was lower. These expression patterns were confirmed by RT-PCR and/or Northern analyses. We compared the expression of some of these genes in the mouse mammary gland under various physiological conditions. The expression levels of genes (clusterin, CBP, and M6P-R) up-regulated in apoptotic conditions were higher at involution than during lactation. On the other hand, genes (Pin, CDC2) downregulated in apoptotic conditions were relatively highly expressed in virgin and pregnant mice. We conclude that certain genes such as clusterin, sFRP-1, GAS1 and CBP are induced in apoptotic mammary epithelial cells, and others are repressed. Moreover, the apoptosis array is an efficient technique for comparing gene expression profiles in different states of the same cell type.

Keywords

Acknowledgement

Supported by : Korea Research Foundation

References

  1. Andoh, T., Chock, P. B., and Chiueh, C. C. (2002) The roles of thioredoxin in protection against oxidative stress-induced apoptosis in SH-SY5Y cells. J. Biol. Chem. 277, 9655-9660 https://doi.org/10.1074/jbc.M110701200
  2. Arnold, N. B., Ketterer, K., Kleeff, J., Friess, H., Buchler, M. W., et al. (2004) Thioredoxin is downstream of Smad7 in a pathway that promotes growth and suppresses cisplatininduced apoptosis in pancreatic cancer. Cancer Res. 64, 3599-3606 https://doi.org/10.1158/0008-5472.CAN-03-2999
  3. Ball, R. K., Friis, R. R., Schonenberger, C.-A., Doppler, W., and Groner, B. (1988) Prolactin regulation of $\beta$-casein gene expression and of a cytosolic 120-kd protein in a cloned mouse mammary epithelial cell line. EMBO J. 7, 2089-2095
  4. Chang, Y. W., Jakobi, R., McGinty, A., Foschi, M., Dunn, M. J., et al. (2000) Cyclooxygenase 2 promotes cell survival by stimulation of dynein light chain expression and inhibition of neuronal nitric oxide synthase activity. Mol. Cell. Biol. 20, 8571-8579 https://doi.org/10.1128/MCB.20.22.8571-8579.2000
  5. Chapman, R. S., Lourenco, P. C., Tonner, E., Flint, D. J., Selbert, S., et al. (1999) Suppression of epithelial apoptosis and delayed mammary gland involution in mice with a conditional knockout of Stat3. Genes Dev. 13, 2604-2616 https://doi.org/10.1101/gad.13.19.2604
  6. Chen, D., Li, M., Luo, J., and Gu, W. (2003) Direct interactions between HIF-1 alpha and Mdm2 modulate p53 function. J. Biol. Chem. 278, 13595-13598 https://doi.org/10.1074/jbc.C200694200
  7. Chomozynski, P. and Sacchi, N. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenolchloroform extraction. Anal. Biochem. 162, 156-159
  8. Chrivia, J. C., Kwok, R. P., Lamb, N., Hagiwara, M., Montminy, M. R., et al. (1993) Phosphorylated CREB binds specifically to the nuclear protein CBP. Nature 365, 855-859 https://doi.org/10.1038/365855a0
  9. Del Sal, G., Ruaro, M. E., Philipson, L., and Schneider, C. (1992) The growth arrest-specific gene, GAS1, is involved in growth suppression. Cell 70, 595-607 https://doi.org/10.1016/0092-8674(92)90429-G
  10. Duquesne, F., Florent, M., Roue, G., Troussard, X., and Sola, B. (2001) Ectopic expression of cyclin D1 impairs the proliferation and enhances the apoptosis of a murine lymphoid cell line. Cell Death Differ. 8, 51-62 https://doi.org/10.1038/sj.cdd.4400768
  11. Feng, Z., Marti, A., Jehn, B., Altermatt, H, J., Chicaiza, G., et al. (1995) Glucocorticoid and progesterone inhibit involution and programmed cell death in the mouse mammary gland. J. Cell Biol. 131, 1095-1103 https://doi.org/10.1083/jcb.131.4.1095
  12. Gupta, V. and Patel, R. C. (2002) Proapoptotic protein PACT is expressed at high levels in colonic epithelial cells in mice. Am. J. Physiol. Gastrointest. Liver Physiol. 283, G801-G808
  13. Haendeler, J., Tischler, V., Hoffmann, J., Zeiher, A. M., and Dimmeler, S. (2004) Low doses of reactive oxygen species protect endothelial cells from apoptosis by increasing thioredoxin- 1 expression. FEBS Lett. 577, 427-433 https://doi.org/10.1016/j.febslet.2004.10.041
  14. Han, X. and Amar, S. (2004) Secreted frizzled-related protein 1 (SFRP1) protects fibroblasts from ceramide-induced apoptosis. J. Biol. Chem. 279, 2832-2840 https://doi.org/10.1074/jbc.M308102200
  15. Han, E. K., Begemann, M., Sgambato, A., Soh, J. W., Doki, Y., et al. (1996) Increased expression of cyclin D1 in a murine mammary epithelial cell line induces p27kip1, inhibits growth, and enhances apoptosis. Cell Growth Differ. 7, 699- 710
  16. Jung, D. J., Bong, J. J., and Baik, M. (2004) Extracellular proteinase inhibitor-accelerated apoptosis is associated with B cell activating factor in mammary epithelial cells. Exp. Cell Res. 292, 115-122 https://doi.org/10.1016/j.yexcr.2003.08.004
  17. Katayama, K., Dobashi, Y., Kitagawa, M., Kamekura, S., Kawai, M., et al. (2001) Overexpression of cdk4/cyclin D1 induces apoptosis in PC12 cells in the presence of trophic support. FEBS Lett. 509, 382-388 https://doi.org/10.1016/S0014-5793(01)03200-8
  18. Kim, T. W., Moon, H. B., and Kim, S. J. (2003) Interleukin-10 is up-regulated by prolactin and serum-starvation in cultured mammary epithelial cells. Mol. Cells 16, 168-172
  19. Ko, J., Ryu, K. S., Lee, Y. H., Na, D. S., Kim, Y. S., et al. (2002) Human secreted frizzled-related protein is downregulated and induces apoptosis in human cervical cancer. Exp. Cell Res. 280, 280-287 https://doi.org/10.1006/excr.2002.5649
  20. Kobayashi, K., Hatano, M., Otaki, M., Ogasawara, T., and Tokuhisa, T. (1999) Expression of a murine homologue of the inhibitor of apoptosis protein is related to cell proliferation. Proc. Natl. Acad. Sci. USA 96, 1457-1462
  21. Kohn, K. W. (1999) Molecular interaction map of the mammalian cell cycle control and DNA repair systems. Mol. Biol. Cell. 8, 2703-2734
  22. Krek, W., Ewen, M. E., Shirodkar, S., Arany, Z., Kaelin, W. G., Jr., et al. (1994) Negative regulation of the growthpromoting transcription factor E2F-1 by a stably bound cyclin A-dependent protein kinase. Cell 78, 161–172 https://doi.org/10.1016/0092-8674(94)90582-7
  23. Lacher, M. D., Siegenthaler, A., Jager, R., Yan, X., Hett, S., et al. (2003) Role of DDC-4/sFRP-4, a secreted frizzled-related protein, at the onset of apoptosis in mammary involution. Cell Death Differ. 10, 528-5 https://doi.org/10.1038/sj.cdd.4401197
  24. Lakins, J., Bennett, S. A., Chen, J. H., Arnold, J. M., Morrissey, C., et al. (1998) Clusterin biogenesis is altered during apoptosis in the regressing rat ventral prostate. J. Biol. Chem. 273, 27887-27895 https://doi.org/10.1074/jbc.273.43.27887
  25. Lee, D., Ha, S., Kho, Y., Kim, J., Cho, K., et al. (1999) Induction of mouse Ca2+-sensitive chloride channel 2 gene during involution of mammary gland. Biochem. Biophys. Res. Commun. 264, 933-937 https://doi.org/10.1006/bbrc.1999.1583
  26. Leskov, K. S., Klokov, D. Y., Li, J., Kinsella, T. J., and Boothman, D. A. (2003) Synthesis and functional analyses of nuclear clusterin, a cell death protein. J. Biol. Chem. 278, 11590-11600 https://doi.org/10.1074/jbc.M209233200
  27. Li, M., Liu, X., Robinson, G., Bar-Peled, U., Wagner, K. U., et al. (1997) Mammary-derived signals activated programmed cell death during the first stage of mammary gland involution. Proc. Natl. Acad. Sci. USA 94, 3425-3430
  28. Lu, M., Kwan, T., Yu, C., Chen, F., Freedman, B., et al. (2005) Peroxisome proliferator-activated receptor gamma agonists promote TRAIL-induced apoptosis by reducing survivin levels via cyclin D3 repression and cell cycle arrest. J. Biol. Chem. 280, 6742-6751 https://doi.org/10.1074/jbc.M411519200
  29. Ludwig, T., Tenscher, K., Remmler, J., Hoflack, B., and Lobel, P. (1994) Cloning and sequencing of cDNAs encoding the full-length mouse mannose 6-phosphate/insulin-like growth factor II. Gene 142, 311-312 https://doi.org/10.1016/0378-1119(94)90282-8
  30. Lund, L. R., Romer, J., Thomasset, N., Solberg, H., Pyke, C., et al. (1996) Two distinct phases of apoptosis in mammary gland involution: proteinase-independent and dependent pathways. Development 122, 181-193
  31. Magee, T. R., Ferrini, M. G., Davila, H. H., Zeller, C. B., Vernet, D., et al. (2003) Protein inhibitor of nitric oxide synthase (NOS) and the N-methyl-D-aspartate receptor are expressed in the rat and mouse penile nerves and colocalize with penile neuronal NOS. Biol. Reprod. 68, 478-488 https://doi.org/10.1095/biolreprod.102.007310
  32. Marti, A., Feng, Z., Altermatt, H. J., and Jaggi, R. (1997) Milk accumulation triggers apoptosis of mammary epithelial cells. Eur. J. Cell. Biol. 73, 158-165
  33. Marti, A., Lazar, H., Ritter, P., and Jaggi, R. (1999) Transcription factor activities and gene expression during mouse mammary gland involution. J. Mammary Gland Biol. Neoplasia 4, 145-152 https://doi.org/10.1023/A:1018721107061
  34. Merlo, G. R., Basolo, F., Fiore, L., Duboc, L., and Hynes, N. E. (1995) p53-dependent and p53-independent activation of apoptosis in mammary epithelial cells reveals a survival function of EGF and insulin. J. Cell Biol. 128, 1185-1196 https://doi.org/10.1083/jcb.128.6.1185
  35. Motyka, B., Korbutt, G., Pinkoski, M. J., Heibein, J. A., Caputo, A., et al. (2000) Mannose 6-phosphate/insulin-like growth factor II receptor is a death receptor for granzyme B during cytotoxic T cell-induced apoptosis. Cell 103, 491-500 https://doi.org/10.1016/S0092-8674(00)00140-9
  36. Rattner, A., Hsieh, J. C., Smallwood, P. M., Gilbert, D. J., Copeland, N. G., et al. (1997) A family of secreted proteins contains homology to the cysteine-rich ligand-binding domain of frizzled receptors. Proc. Natl. Acad. Sci. USA 94, 2859-2863
  37. Ravnik, S. E. and Wolgemuth, D. J. (1996) The developmentally restricted pattern of expression in the male germ line of a murine cyclin A, cyclin A2, suggests roles in both mitotic and meiotic cell cycles. Dev. Biol. 173, 69-78 https://doi.org/10.1006/dbio.1996.0007
  38. Rowlands, T. M., Pechenkina, I. V., Hatsell, S. J., Pestell, R. G., and Cowin, P. (2003) Dissecting the roles of beta-catenin and cyclin D1 during mammary development and neoplasia. Proc. Natl. Acad. Sci. USA 100, 11400-11405
  39. Ruaro, E. M., Collavin, L., Del Sal, G., Haffner, R., Oren, M., et al. (1997) A proline-rich motif in p53 is required for transactivation- independent growth arrest as induced by GAS1. Proc. Natl. Acad. Sci. USA 94, 4675-4680
  40. Simons, A., Melamed-Bessudo, C., Wolkowicz, R., Sperling, J., Sperling, R., et al. (1997) PACT: cloning and characterization of a cellular p53 binding protein that interacts with Rb. Oncogene 14, 145-155 https://doi.org/10.1038/sj.onc.1200825
  41. Strange, R., Li, F., Saurer, S., Burkhardt, A., and Friis, R. R. (1992) Apoptotic cell death and tissue remodelling during mouse mammary gland involution. Development 115, 49-58
  42. Talhouk, R. S., Bissell, M. J., and Werb, Z. (1992) Coordinated expression of extracellular matrix-degrading proteinases and their inhibitors regulates mammary epithelial function during involution. J. Cell Biol. 118, 1271-1282 https://doi.org/10.1083/jcb.118.5.1271
  43. Tokunaga, K., Taniguchi, H., Yoda, K., Shimizu, M., and Sakiyama, S. (1986) Nucleotide sequence of a full-length cDNA for mouse cytoskeletal beta-actin mRNA. Nucleic Acids Res. 14, 2829 https://doi.org/10.1093/nar/14.6.2829
  44. Topper, Y. J. and Freeman, C. S. (1980) Multiple hormone interactions in the developmental biology of the mammary gland. Physiol. Rev. 60, 1049-1106
  45. Walker, N. I., Bennett, R. E., and Kerr, J. F. (1989) Cell death by apoptosis during involution of the lactating breast in mice and rats. Am. J. Anat. 185, 19-32 https://doi.org/10.1002/aja.1001850104
  46. Yamamoto, T. and Tanigawa, N. (2001) The role of survivin as a new target of diagnosis and treatment in human cancer. Med. Electron. Microsc. 34, 207-212 https://doi.org/10.1007/s007950100017
  47. Yang, R., Muller, C., Huynh, V., Fung, Y. K., Yee, A. S., et al. (1999) Functions of cyclin A1 in the cell cycle and its interactions with transcription factor E2F-1 and the Rb family of proteins. Mol. Cell Biol. 19, 2400-2407
  48. Zamorano, A., Lamas, M., Vergara, P., Naranjo, J. R., and Segovia, J. (2003) Transcriptionally mediated gene targeting of GAS1 to glioma cells elicits growth arrest and apoptosis. J. Neurosci. Res. 71, 256-263 https://doi.org/10.1002/jnr.10461