DOI QR코드

DOI QR Code

Differential Proteome Expression of in vitro Proliferating Hanwoo Stromal Vascular Cells from Omental, Subcutaneous and Intramuscular Depots in Response to Hormone Deprivation and IGF-1, Estradiol-17β Addition

  • Rajesh, Ramanna Valmiki (Division of Animal Genomics and Bioinformatics, National Institute of Animal Science, Rural Development Administration) ;
  • Kim, Seong-Kon (Division of Animal Genomics and Bioinformatics, National Institute of Animal Science, Rural Development Administration) ;
  • Park, Min-Ah (Department of Agricultural Science, Korea National Open University) ;
  • Kwon, Seulemina (Division of Animal Genomics and Bioinformatics, National Institute of Animal Science, Rural Development Administration) ;
  • Chang, Jong-Soo (Department of Agricultural Science, Korea National Open University) ;
  • Yoon, Du-Hak (Division of Animal Genomics and Bioinformatics, National Institute of Animal Science, Rural Development Administration) ;
  • Kim, Tae-Hun (Division of Animal Genomics and Bioinformatics, National Institute of Animal Science, Rural Development Administration) ;
  • Lee, Hyun-Jeong (Division of Animal Genomics and Bioinformatics, National Institute of Animal Science, Rural Development Administration)
  • Received : 2010.04.07
  • Accepted : 2010.06.09
  • Published : 2010.06.30

Abstract

The aim of this study was to analyze the proteome expressions of proliferating stromal vascular cells from Hanwoo omental, subcutaneous and intramuscular depots subjected to hormone deprivation and IGF-1, Estradiol-$17{\beta}$ addition. For hormone deprivation or addition studies, the cells were either grown in 10% charcoal-dextran stripped fetal bovine serum (CD-FBS) or in 10% FBS supplemented medium. Further, to analyze the effect of insulin like growth factor (IGF-1) and $17\beta$-Estradiol (E2), cells were grown in 10% CD-FBS containing IGF-1 (10 ng/ml) or E2 (10 nM). The results showed that hormone deprivation had a negative impact on proliferation among the cells from all depots without any growth difference. On comparison of proliferation levels, higher levels were observed in cells that were grown in 10% FBS than in 10% CD-FBS alone or with IGF-1/E2. Proteome expression from preadipocytes grown in hormone deprivation conditions were compared by 2D-DIGE and MALDIToF/ToF. A total of twelve different proteins were found to be differentially expressed under hormone deprivation conditions. Further, our proteomic analysis with DIGE under IGF-1 and E2 addition revealed four proteins with differential expression levels. Moreover, the results highlighted in this study offer a role for each differentially expressed protein with respect to their effect in positive or negative regulation on proliferation.

Keywords

References

  1. Baker, S. J. 2003. Small unstable apoptotic protein, an apoptosisassociated protein, suppresses proliferation of myeloid cells. Cancer Res. 63:705-712.
  2. Belanger, C., Luu-The, V., Dupont, P. and Tchernof, A. 2002. Adipose tissue intracrinology: potential importance of local androgen/estrogen metabolism in the regulation of adiposity. Horm. Metab. Res. 34:737-745. https://doi.org/10.1055/s-2002-38265
  3. Blishchenko, E. Y., Sazonova, O. V., Kalinina, O. A., Yatskin, O. N., Philippova, M. M., Surovoy, A. Y., Karelin, A. A. and Ivanov, V. T. 2002. Family of hemorphins: co-relations between aminoacid sequences and effects in cell cultures. Peptides 23:903-910. https://doi.org/10.1016/S0196-9781(02)00017-7
  4. Brook, C. G., Llyod, J. K. and Wolf, O. H. 1972. Relation between age of onset of obesity and size and number of adipose cells. BMJ 2:25-27. https://doi.org/10.1136/bmj.2.5804.25
  5. Byun, Y., Chen, F., Chang, R., Trivedi, M., Green, K. J. and Cryns, V. L. 2001. Caspase cleavage of vimentin disrupts intermediate filaments and promotes apoptosis. Cell Death Differ. 8:443-450. https://doi.org/10.1038/sj.cdd.4400840
  6. Chi, S., Kitanaka, C., Noguchi, K., Mochizuki, T., Nagashima, Y., Shirouzu, M., Fujita, H., Yoshida, M., Chen, W., Asai, A., Himeno, M., Yokoyama, S. and Kuchino, Y. 1999. Oncogenic Ras triggers cell suicide through the activation of a caspaseindependent cell death program in human cancer cells. Oncogene 18:2281-2290. https://doi.org/10.1038/sj.onc.1202538
  7. Cianzio, D., Topel, D. G., Whitehurst, G. and Beitz, D. C. 1982. Adipose tissue growth in cattle representing two frame sizes: Distribution among depots. J. Anim. Sci. 55:305-312. https://doi.org/10.2527/jas1982.552305x
  8. Cornelius, P., MacDougald, O. A. and Lane, M. D. 1994. Regulation of adipocyte development. Annu. Rev. Nutr.14:99-129. https://doi.org/10.1146/annurev.nu.14.070194.000531
  9. Cryer, J., Woodhead, B. G. and Cryer, A. 1987. The isolation and characterization of a putative adipocyte precursor cell type from the white adipose tissue of the chicken (Gallus domesticus). Comp. Biochem. Physiol. 86A:515-520.
  10. Deslex, S., Negrel, R. and Ailhaud, G. 1987. Development of a chemically defined serum free medium for differentiation of rat precursor cells. Exp. Cell. Res. 168:15-30. https://doi.org/10.1016/0014-4827(87)90412-5
  11. Dieudonne, M. N., Pecquery, R. and Guidicelli, Y. 1992. Characteristics of the $\alpha$2/$\beta$-adrenoreceptor-coupled adenylate cyclase system and their relationship with adrenergic responsiveness in hamster fat cells from different anatomical sites. Eur. J. Biochem. 205:867-873. https://doi.org/10.1111/j.1432-1033.1992.tb16851.x
  12. Dieudonne, M. N., Pecquery, R., Leneveu, M. C. and Giudicelli, Y. 2000. Opposite effects of androgens and estrogens on adipogenesis in rat preadipocytes: evidence for sex and siterelated specificities and possible involvement of insulin-like growth factor 1 receptor and peroxisome proliferator-activated receptor gamma2. Endocrinology 141:649-656. https://doi.org/10.1210/en.141.2.649
  13. Dreuillet, C., Tillit, J., Kress, M. and Ernoult-Lange, M. 2002. In vivo and in vitro interaction between human transcription factor MOK2 and nuclear lamin A/C. Nucleic Acids Res. 30:4634-4642. https://doi.org/10.1093/nar/gkf587
  14. Dubey, R. K., Gillespie, D. G., Mi, Z., Rosselli, M., Keller, P. J. and Jackson, E. K. 2000. Estradiol inhibits smooth muscle cell growth in part by activating the cAMP-adenosine pathway. Hypertension 35:262-266. https://doi.org/10.1161/01.HYP.35.1.262
  15. Eskes, R., Desagher, S., Antonsson, B. and Martinou, J. C. 2000. Bid induces the oligomerization and insertion of bax into the outer mitochondrial membrane. Mol. Cell. Biol. 20:929-935. https://doi.org/10.1128/MCB.20.3.929-935.2000
  16. Fajas, L. 2003. Adipogenesis: a cross-talk between cell proliferation and cell differentiation. Ann. Med. 35:79-85. https://doi.org/10.1080/07853890310009999
  17. Finaly, G. A., York, B., Karas, R. H., Fanburg, B. L., Zhang, H., Kwiatkowski, D. J. and J. Noonan, D. 2004. Estrogen-induced smooth muscle cell growth is regulated by tuberin and associated with altered activation of platelet-derived growth factor receptor-$\beta$ and ERK-1/2. J. Biol. Chem. 279:23114-23122. https://doi.org/10.1074/jbc.M401912200
  18. Ghosh, S., Strum, J. C., Sciorra, V. A., Daniel, L. and Bell, R. M. 1996. Raf-1 Kinase possesses distinct binding domains for phosphatidylserine and phosphatidic acid. J. Biol. Chem. 271: 8472-8480. https://doi.org/10.1074/jbc.271.14.8472
  19. Grunewald, T. G. P., Kammerer, U., Wrinkler, C., Schindler, D., Sickmann, A,. Honig, A. and Butt, E. 2007. Overexpression of LASP-1 mediates migration and proliferation of human ovarian cancer cells and influence zyxin localization. Br. J. Cancer 96:296-305. https://doi.org/10.1038/sj.bjc.6603545
  20. Hauner, H., Schmid, P. and Pfeiffer, E. F. 1987. Glucocorticoids and insulin promote the differentiation of human adipocyte precursor cells into fat cells. J. Clin. Endocrinol. Metab. 64: 832-835. https://doi.org/10.1210/jcem-64-4-832
  21. Hausman, D. B., DiGirolamo, M., Bartness, T. J. and Hausman, G. J. 2001. The biology of white adipocyte proliferatio. Obes Rev 2:239-254. https://doi.org/10.1046/j.1467-789X.2001.00042.x
  22. Hausman, G. J. and Richardson, R. L. 1998. Newly recruited and preexisting preadipocytes in cultures of porcine stromalvascular cells: morphology, expression of extracellular matrix components, and lipid accretion. J. Anim. Sci. 76:48-60. https://doi.org/10.2527/1998.76148x
  23. Hausman, G. J., Wright, J. T., Dean, R. and Richardson, R. L. 1993. Cellular and molecular aspects of the regulation of adipogenesis. J. Anim. Sci. 71:33-55.
  24. Hellmann, U., Wemstedt, C., Gonez, J. and Heldin, C. H. 1995. Improvement of an “In-gel” digestion procedure for the micropreparation of internal protein fragments for amino acid sequencing. Anal. Biochem. 224:451-455. https://doi.org/10.1006/abio.1995.1070
  25. Hoffrogge, R., Mikkat, S., Scharf, C., Beyer, S., Christoph, H., Pahnke, J., Mix, E., Berth, M., Uhrmacher, A., Zubrzycki, I. Z., Miljan, E., Volker, U. and Rolfs, A. 2006. 2-DE proteome analysis of a proliferating and differentiating human neuronal stem cell line (ReNcell VM). Proteomics 6:1833-1847. https://doi.org/10.1002/pmic.200500556
  26. Horiuchi, A., Nikaido, T., Taniguchi, S. and Fuji, S. 1999. Possible role of calponin h1 as a tumor suppressor in human uterine liomyosarcoma. J. Natl. Cancer Inst. 91:790-795. https://doi.org/10.1093/jnci/91.9.790
  27. Hughes, B. W., King, S. A., Allan, P. W., Parker, W. B. and Sorscher, E. J. 1998. Cell to cell contact is not required for bystander cell killing by Escherichia coli purine nucleoside phosphorylase. J. Biol. Chem. 273:2322-2328. https://doi.org/10.1074/jbc.273.4.2322
  28. Hughes, B. W., Wells, A. H., Bebok, Z., Gadi, V. K., Jr. Garver, R. I., Parker, W. B. and Sorscher, E. J. 1995. Bystander killing of melanoma cells using the human tyrosinase promoter to express the Escherichia coli purine nucleoside phosphorylase gene. Cancer Res. 55:3339-3345.
  29. Hung, Y., Jin, Y., Yan, C., Yu, Y., Bai, J., Chen, F., Zhao, Y. and Fu, S. 2008. Involvement of Annexin A2 in p53 induced apoptosis in lung cancer. Mol. Cell. Biochem. 309:117-123. https://doi.org/10.1007/s11010-007-9649-5
  30. Iwanami, N., Higuchi, T., Sasano, Y., Fujiwara, T., Hoa, V. Q., Okada, M., Talukder, S. R., Kunimatsu, S., Li, J., Saito, F., Bhattacharya, C., Matin, A., Sasaki, T., Shimizu, N., Mitani, H., Himmelbauer, H., Momoi, A., Kondoh, H., Furutani-Seiki, M. and Takahama, Y. 2008. WDR55 is a mucleolar modulator of ribosomal RNA synthesis, cell cycle progression, and Teleost organ development. PLoS Genet. 4:1-13. https://doi.org/10.1371/journal.pgen.0040001
  31. Jae, H. H., Heo, J. S. and Lee, Y. J. 2006. Estradiol-17$\beta$ stimulates proliferation of mouse embryonic stem cells: involvement of MAPKs and CDKs as well as protooncogenes, Am. J. Physiol., Cell Physiol. 290:C1067-1075. https://doi.org/10.1152/ajpcell.00222.2005
  32. Kampa, M., Bakogeorgou, E., Hatzolgou, A., Damianaki, A., Martin, P. M. and Castanas, E. 1997. Opioid alkaloids and casomorphin peptides decrease the proliferation of prostatic cancer cell lines (LNCaP, PC3 and DU145) through a partial interaction with opioid receptors. Eur. J. Pharmacol. 335:255-265. https://doi.org/10.1016/S0014-2999(97)01213-2
  33. Kawakami, A., Nakashima, T., Tsuboi, M., Urayama, S., Matsuoka, N., Ida, H., Kawabe, Y., Sakai, H., Migita, K., Aoyagi, T., Nakashima, M., Maeda, K. and Eguchi, K. 1998. Insulin-like growth factor I stimulates proliferation and Fas-mediated apoptosis of human osteoblasts. Biochem. Biophys. Res. Commun. 247:46-51. https://doi.org/10.1006/bbrc.1998.8728
  34. Kirkland, J. and Harris, P. M. 1980. Changes in adipose tissue of the rat due to early under nutrition followed by rehabilitation. 3. Changes in cell replication studies with tritiated thymidine. Br. J. Nutr. 43:33-43. https://doi.org/10.1079/BJN19800062
  35. Kras, K. M., Hausman, D. B., Hausman, G. J. and Martin, R. J. 1999. Adipocyte development is dependent upon stem cell recruitment and proliferation of preadipocytes. Obes. Res. 7:491-497. https://doi.org/10.1002/j.1550-8528.1999.tb00438.x
  36. Krotkiewski, M., Bjorntorp, P., Sjostrom, L. and Smith. U. 1983. Impact of obesity on metabolism in men and women. Importance of regional adipose tissue distribution. J. Clin. Invest. 72: 1150-1162. https://doi.org/10.1172/JCI111040
  37. Lal, S., Lauer, U. M., Niethammer, D., Beck, J. F. and Schlegel, P. G. 2000. Suicide genes: past, present and future perspectives. Immunol. Today 21:48-54 https://doi.org/10.1016/S0167-5699(99)01550-9
  38. Landry, F., Lombardo, C. R. and Smith, J. W. 2000. A method for application of samples to Matrix.Assisted Laser Desorption Ionization Time-of-Flight targets that enhances peptide detection. Anal. Biochem. 279:1-8. https://doi.org/10.1006/abio.1999.4468
  39. Li, D. and Roberts, R. 2001. WD-repeat proteins: structure characteristics, biological function, and their involvement in human diseases. Cell. Mol. Life Sci. 58:2085-2097. https://doi.org/10.1007/PL00000838
  40. Liao, X., Li, S., Lou, Z., Chen, Z. and Bartlam, M. 2007. Crystallization and Preliminary Crystallographic Analysis of Transgelin. Protein Pept. Lett. 14:209-211. https://doi.org/10.2174/092986607779816122
  41. Lindquist, D. L. and de Alarcon, P. A. 1987. Charcoal-dextran treatment of fetal bovine serum removes an inhibitor of human CFU-megakaryocytes. Exp. Hematol. 15:234-238.
  42. Lloyd, D. J., Trembath, R. C. and Shackleton, S. 2002. A novel interaction between lamin A and SREBP1: implications for partial lipodystrophy and other laminopathies. Hum. Mol. Genet. 11:769-777. https://doi.org/10.1093/hmg/11.7.769
  43. Mancini, M., Nicholson, D. W., Roy, S., Thornberry, N. A., Peterson, E. P., Casciola-Rosen, L. A. and Rosen, A. 1998. The caspase-3 precursor has a cytosolic and mitochondrial distribution: Implications for apoptotic signaling. J. Cell. Biol. 140:1485-1495. https://doi.org/10.1083/jcb.140.6.1485
  44. Niesler, C. U., Siddle, K. and Prins, J. B. 1998. Human preadipocytes display susceptibility to apoptosis. Diabetes 47: 1365-1368. https://doi.org/10.2337/diabetes.47.8.1365
  45. Niesler, C. U., Urso, B., Prins, J. B. and Siddle, K. 2000. IGF-1 inhibits apoptosis induced by serum withdrawal, but potentiates TNF-$\alpha$-induced apoptosis, in 3T3-L1 preadipocytes. J. Endocrinol. 167:165-174. https://doi.org/10.1677/joe.0.1670165
  46. Nishimura, T., Hattori, A. and Takahashi, K. 1999. Structural changes in intramuscular connective tissue during the fattening of Japanese black cattle: Effect of marbling on beef tenderization. J. Anim. Sci. 77:93-104 https://doi.org/10.2527/1999.77193x
  47. Papineau, D., Gagnon, A. and Sorisky, A. 2003. Apoptosis of human abdominal preadipocytes before and after differentiation into adipocytes in culture. Metab. Clin. Exp. 52:987-992 https://doi.org/10.1016/S0026-0495(03)00165-3
  48. Park, S. H., Yoon, B. C. and Han, H. J. 2006. The effect of insulin-like grow factor-1 on cell proliferation and its related signal pathways in primary cultured hepatocytes. Lab. Anim. Res. 22:215-219.
  49. Parker, W. B., Allan, P. W., Shaddix, S. C., Rose, L. M., Speegle, H. F., Gillespie, G. Y. and Jr. Bennett, L. L. 1998. Metabolism and metabolic actions of 6-methylpurine and 2-fluoroadenine in human cells. Biochem. Pharmacol. 55:1673-1681. https://doi.org/10.1016/S0006-2952(98)00034-3
  50. Pedersen, S. B., Fuglsig, S., Sjogren, P. and Richelsen, B. 1996. Identification of steroid receptors in human adipose tissue. Eur. J. Clin. Invest. 26:1051-1056. https://doi.org/10.1046/j.1365-2362.1996.380603.x
  51. Penuelas, S., Noe, V. and Ciudad, C. J. 2005. Modulation of IMPDH2, surviving, topoisomerase I and vimentin increases sensitivity to methotrexate in HT29 human colon cancer cells. FEBS J. 272:696-710. https://doi.org/10.1111/j.1742-4658.2004.04504.x
  52. Pico, C., Puigserver, P., Oliver, P. and Palou, A. 1998. 2-Methoxyestradiol, an endogenous metabolite of 17$\beta$-estradiol, inhibits adipocyte proliferation. Mol. Cell. Biochem. 189:1-7. https://doi.org/10.1023/A:1006835124149
  53. Pietras, R. J., Nemere, I. and Szego, C. M. 2001. Steroid hormone receptors in target cell membranes, Endocrine 14:417-427. https://doi.org/10.1385/ENDO:14:3:417
  54. Prins, J. B. and O'Rahilly, S. 1997. Regulation of adipose cell number in man. Clin Sci 92:3-11. https://doi.org/10.1042/cs0920003
  55. Rabilloud, T. 1998. Use of thiourea to increase the solubility of membrane proteins in two-dimensional electrophoresis. Electrophoresis 19:758-760. https://doi.org/10.1002/elps.1150190526
  56. Rebuffe-Scrive, M., Enk, L. and Crona, N. 1985. Fat cell metabolism in differentregions in women. Effect of menstrual cycle, pregnancy and lactation. J. Clin. Invest. 75:1973-1976. https://doi.org/10.1172/JCI111914
  57. Rescher, U. and Gerke, V. 2004. Annexins-unique membrane binding proteins with diverse functions. J. Cell. Sci. 117:2631-2639. https://doi.org/10.1242/jcs.01245
  58. Rodriguez-Cuenca, S., Monjo, M., Proenza, A. M. and Roca, P. 2005. Depot differences in steroid receptor expression in adipose tissue: possible role of the local steroid milieu. Am. J. Physiol. Endocrinol. Metab. 288:E200-207. https://doi.org/10.1152/ajpendo.00270.2004
  59. Roncari, D. A. and Van, R. L. 1978. Promotion of human adipocyte precursor replication by 17$\beta$estradiol in culture. J. Clin. Invest. 62:503-508 https://doi.org/10.1172/JCI109153
  60. Rong, W., Jian, D., Zhen-Ming, Z., Li-Ping, R. and Qing-Xiang. M. 2007. Isolation and in vitro culture of intramuscular preadipocytes from Luxi adult yellow cattle. Chin. J. Agric. Biotechol. 4:229-232
  61. Sewing, A., Wiseman, B., Lloyd, A. C. and Land, H. 1997. Highintensity Raf signal causes cell cycle arrest mediated by p21Cip1. Mol. Cell. Biol. 17:5588-5597. https://doi.org/10.1128/MCB.17.9.5588
  62. Simpson, E., Rubin, G., Clyne, C., Robertson, K., O'Donnell, L., Jones, M. and. Davis, S. 2000. The role of local estrogen biosynthesis in males and females. Trends Endocrinol. Metab. 11:184-188. https://doi.org/10.1016/S1043-2760(00)00254-X
  63. Smyth, M. J., Sparks, R. L. and Wharton, W. 1993. Preadipocyte cell lines: models of cellular proliferation and differentiation. J. Cell. Sci. 106:1-9.
  64. Sommers, C. L., Byers, S. W., Thompson, E. W., Torri, J. A. and Gelmann, E. P. 1994. Differentiation state and invasiveness of human breast cancer cell lines. Breast Cancer Res. Treat. 31:325-335. https://doi.org/10.1007/BF00666165
  65. Sorscher, E. J., Peng, S., Bebok, Z., Allan, P. W., Bennett Jr. L. L. and Parker, W. B. 1994. Tumor cell bystander killing in colonic carcinoma utilizing the Escherichia coli DeoD gene to generate toxic purines. Gene Ther. 1:233-238.
  66. Spaulding, H. L., Saijo, F., Turnage, R. H., Alexander, J. S., Aw, T. Y. and Kalogeris, T. J. 2006. Apolipoprotein A-IV attenuates oxidant induced apoptosis in mitotic competent, undifferentiated cells by modulating intracellular glutathione redox balance. Am. J. Physiol., Cell Physiol. 290:C95-103. https://doi.org/10.1152/ajpcell.00388.2005
  67. Takahashi, K., Miwa, A., Tkagi, M., Kojima, A., Eshita, Y. and Nara, Y. 1995. Overexpression of calponin in transgenic rats inhibits intimal arterial smooth muscle cell hyperplasia in the carotid artery model of restenosis. Circulation 92:1-295. https://doi.org/10.1161/01.CIR.92.9.1
  68. Takahashi, K., Tkagi, M., Ohgami, K., Nakai, M., Kojima, A. and Nadal-Ginard, B. 1993. Inhibition of smooth muscle cell migration and proliferation caused by transfection of the human calponin gene is associated with enhanced cell matrix adhesion and reduced PDGF responsiveness. Circulation 88:1-74.
  69. Tong, W. and Pollard, J. W. 2002. The Endometrium. (Glasser, S. R., Aplin, J. D., Giudice, L. C. and Tabibzadeh, S. (eds), Taylor and Francis, London, pp. 94-109.
  70. Tonge, R., Shaw, J., Middleton, B., Rowlinson, R., Rayner, S., Young, J., Pognan, F., Hawkins, E., Currie, I. and Davison, M. 2001. Validation and development of fluorescence twodimensional differential gel electrophoresis proteomics technology. Proteomics1:377-396. https://doi.org/10.1002/1615-9861(200103)1:3<377::AID-PROT377>3.0.CO;2-6
  71. Van de Klundert, F. A., Jansen, H. J. and Bloemendal, H. 1995. Negative regulation of a special, double AP-1 consensus element in the vimentin promoter: interference by the retinoic receptor. J. Cell. Physiol. 164:95-92.
  72. Wei, M. C., Lindsten, T., Mootha, V. K., Weiler, S., Gross, A., Ashiya, M., Thompson, C. B. and Korsmeyer, S. J. 2000. tBID, a membrane-targeted death ligand, oligomerizes BAK to release cytochrome c. Genes Dev. 14:2060-2071.
  73. Wright, J. T. and Hausman, G. J. 1995. Insulin like growth factor-1 (IGF-1) induced stimulation of porcine preadipocyte replication. In Vitro Cell. Dev. Biol. 31:404-408. https://doi.org/10.1007/BF02634290
  74. Xu, Y., Nguyen, Q., Lo, D. C. and Czaja, M. J. 1997. c-mycdependent hepatoma cell apoptosis results from oxidative stress and not a deficiency of growth factors. J. Cell. Physiol. 170:192-199. https://doi.org/10.1002/(SICI)1097-4652(199702)170:2<192::AID-JCP11>3.0.CO;2-K
  75. Yang, B. C., Chang, H. M., Wang, Y. S., Chen, R. F. and Lin, S. J. 1996. Transient induction of apoptosis in serum-starved glioma cells by insulin and IGF-I. Biochim. Biophys. Acta. 1314:83-92. https://doi.org/10.1016/S0167-4889(96)00079-1
  76. Zhu, J., Woods, D., McMahon, M. and Bishop, J. M. 1998. Senescence of human fibroblasts induced by oncogenic Raf. Genes Dev. 12:2997-2307. https://doi.org/10.1101/gad.12.19.2997