L. Kovalevska, A. Matveeva, O. Kashuba

RE Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology of National Academy of Sciences of Ukraine


Summary. The present paper is a review on the literature data and our research, concerning a role of the retinoblastoma (RB) protein in pluripotent stem cells (SCs) proliferation and differentiation. The role of RB in the cell cycle, in development of pathological conditions, in the control on cell differentiation, and in epigenetic regulation of expression of the SC-specific genes is discussed, in the light of protein-protein interactions. The present review might be useful for the further studies and understanding of SC properties, as well as the effectiveness of use of SCs in medical practice.

Keywords: RB protein, RB-binding proteins, stemness, pluripotent cells, cell cycle, transcription factors.

DOI: https://doi.org/10.15407/oncology.2023.04.245


  1. Hanahan D, Weinberg RA. The hallmarks of cancer. Cell 2000; 100(1): 57–70.
  2. Weinberg RA. The Biology of Cancer. Jeffcock E (eds). USA: Garland Science, Taylor and Francis Group; 2007: 255–300.
  3. Zhu L. Tumour suppressor retinoblastoma protein Rb: a transcriptional regulator. Eur J Cancer 2005; 41 (16): 2415–27. doi: 10.1016/j.ejca.2005.08.009.
  4. Giacinti C, Giordano A. RB and cell cycle progression. Oncogene 2006; 25 (38): 5220–7. doi: 10.1038/sj.onc.1209615.
  5. Rodriguez A, Kelley C, Patel A, Ramasubramanian A. Prenatal diagnosis of retinoblastomas: a scoping review. Int J Gen Med 2023; 16: 1101–10. doi: 10.2147/IJGM.S380634.
  6. Figueiredo D, Marques IA, Pires AS, et al. Risk of second tumors in retinoblastoma survivors after ionizing radiation: a review. Cancers (Basel) 2023; 15 (22): 5336. doi: 10.3390/cancers15225336.
  7. Toguchida J, McGee TL, Paterson JC, et al. Complete genomic sequence of the human retinoblastoma susceptibility gene. Genomics 1993; 17 (3): 535–43. doi: 10.1006/geno.1993.1368.
  8. Goodrich DW. The retinoblastoma tumor-suppressor gene, the exception that proves the rule. Oncogene 2006; 25 (38): 5233–43. doi: 10.1038/sj.onc.1209616.
  9. Sherr CJ. Mammalian G1 cyclins. Cell 1993; 73 (6): 1059–65. doi: 10.1016/0092-8674(93)90636-5.
  10. Hunter T, Pines J. Cyclins and cancer. II: Cyclin D and CDK inhibitors come of age. Cell 1994; 79 (4): 573–82. doi: 10.1016/0092-8674(94)90543-6.
  11. Morgan DO. Principles of CDK regulation. Nature 1995; 374 (6518): 131–4. doi: 10.1038/374131a0.
  12. Hirama T, Koeffler HP. Role of the cyclin-dependent kinase inhibitors in the development of cancer. Blood 1995; 86 (3): 841–54.
  13. Schwaller J, Pabst T, Koeffler HP, et al. Expression and regulation of G1 cell-cycle inhibitors (p16INK4A, p15INK4B, p18INK4C, p19INK4D) in human acute myeloid leukemia and normal myeloid cells. Leukemia 1997; 11 (1): 54–63. doi: 10.1038/sj.leu.2400522.
  14. Conklin JF, Baker J, Sage J. The RB family is required for the self-renewal and survival of human embryonic stem cells. Nature communications 2012; 3(1244): 1–12. doi: 10.1038/ncomms2254.
  15. Giraddi RR, Shehata M, Gallardo M, et al. Stem and progenitor cell division kinetics during postnatal mouse mammary gland development. Nature communications 2015; 6:8487. doi: 10.1038/ncomms9487.
  16. Becker KA, Ghule PN, Therrien JA, et al. Self-renewal of human embryonic stem cells is supported by a shortened G1 cell cycle phase. Journal of cellular physiology 2006; 209 (3): 883–93. doi: 10.1002/jcp.20776.
  17. Varjosalo M, Keskitalo S, Van Drogen A, et al. The protein interaction landscape of the human CMGC kinase group. Cell reports 2013; 3 (4): 1306–20. doi: 10.1016/j.celrep.2013.03.027.
  18. Ferron SR, Pozo N, Laguna A, et al. Regulated segregation of kinase Dyrk1A during asymmetric neural stem cell division is critical for EGFR-mediated biased signaling. Cell Stem Cell 2010; 7 (3): 367–79. doi: 10.1016/j.stem.2010.06.021.
  19. Kashuba E, Yurchenko M, Yenamandra SP, et al. EBV-encoded EBNA-6 binds and targets MRS18-2 to the nucleus, resulting in the disruption of pRb-E2F1 complexes. Proc Natl Acad Sci U S A 2008; 105 (14): 5489–94. doi: 10.1073/pnas.0801053105.
  20. Kashuba E, Yenamandra PS, Darekar SD, et al. MRPS18-2 protein immortalizes primary rat embryonic fibroblasts and endows them with stem cell-like properties. Proc Natl Acad Sci U S A 2009; 106 (47): 19866–71. doi: 10.1073/pnas.0911545106.
  21. Yenamandra SP, Darekar SD, Kashuba V, et al. Stem cell gene expression in MRPS18-2-immortalized rat embryonic fibroblasts. Cell Death Dis 2012; 19 (3): 138. doi: 10.1038/cddis.2011.138.
  22. Gagrica S, Hauser S, Kolfschoten I, et al. Inhibition of oncogenic transformation by mammalian Lin-9, a pRB-associated protein. The EMBO journal 2004; 23 (23): 4627–38. doi: 10.1038/sj.emboj.7600470.
  23. Esterlechner J, Reichert N, Iltzsche F, et al. LIN9, a subunit of the DREAM complex, regulates mitotic gene expression and proliferation of embryonic stem cells. PLoS One 2013; 8 (5): e62882. doi: 10.1371/journal.pone.0062882.
  24. Mushtaq M, Gaza HV, Kashuba EV. Role of the RB-Interacting Proteins in Stem Cell Biology. Adv Cancer Res 2016; 131: 133–57. doi: 10.1016/bs.acr.2016.04.002.
  25. MacLellan WR, Xiao G, Abdellatif M, et al. A novel Rb- and p300-binding protein inhibits transactivation by MyoD. Molecular and cellular biology 2000; 20 (23): 8903–15. doi: 10.1128/MCB.20.23.8903-8915.2000.
  26. Fang F, Xu Y, Chew KK, et al. Coactivators p300 and CBP maintain the identity of mouse embryonic stem cells by mediating long-range chromatin structure. Stem Cells 2014; 32 (7): 1805–16. doi: 10.1002/stem.1705.
  27. Vitelli L, Condorelli G, Lulli V, et al. A pentamer transcriptional complex including tal-1 and retinoblastoma protein downmodulates c-kit expression in normal erythroblasts. Molecular and cellular biology 2000; 20(14): 5330–42. doi: 10.1128/MCB.20.14.5330-5342.2000.
  28. Welinder E, Murre C. Ldb1, a new guardian of hematopoietic stem cell maintenance: Nat Immunol 2011 ; 12 (2): 113–4. doi: 10.1038/ni0211-113.
  29. Li L, Jothi R, Cui K, et al. Nuclear adaptor Ldb1 regulates a transcriptional program essential for the maintenance of hematopoietic stem cells. Nat Immunol 2011; 12 (2): 129–36. doi: 10.1038/ni.1978.
  30. Yi F, Pereira L, Hoffman JA, et al. Opposing effects of Tcf3 and Tcf1 control Wnt stimulation of embryonic stem cell self-renewal. Nature cell biology 2011; 13 (7): 762–70. doi: 10.1038/ncb2283.
  31. Nam CH, Rabbitts TH. The role of LMO2 in development and in T cell leukemia after chromosomal translocation or retroviral insertion. Mol Ther 2006; 13 (1): 15–25. doi: 10.1016/j.ymthe.2005.09.010.
  32. Warren AJ, Colledge WH, Carlton MB, et al. The oncogenic cysteine-rich LIM domain protein rbtn2 is essential for erythroid development. Cell 1994; 78 (1): 45–57. doi: 10.1016/0092-8674(94)90571-1.
  33. Gering M, Yamada Y, Rabbitts TH, et al. Lmo2 and Scl/Tal1 convert non-axial mesoderm into haemangioblasts which differentiate into endothelial cells in the absence of Gata1. Development 2003; 130 (25): 6187–99. doi: 10.1242/dev.00875.
  34. Pichler G, Wolf P, Schmidt CS, et al. Cooperative DNA and histone binding by Uhrf2 links the two major repressive epigenetic pathways. J Cell Biochem 2011; 112 (9): 2585–93. doi: 10.1002/jcb.23185.
  35. Tokitou F, Nomura T, Khan MM, et al. Viral ski inhibits retinoblastoma protein (Rb)-mediated transcriptional repression in a dominant negative fashion. The Journal of biological chemistry 1999; 274 (8): 4485–8. doi: 10.1074/jbc.274.8.4485.
  36. Deheuninck J, Luo K. Ski and–57. doi: 10.1038/cr.2008.324.
  37. Nicol R, Stavnezer E. Transcriptional repression by v-Ski and c-Ski mediated by a specific DNA binding site. The Journal of biological chemistry 1998; 273 (6): 3588–97. doi: 10.1074/jbc.273.6.3588.
  38. Blank U, Karlsson G, Moody JL, et al. Smad7 promotes self-renewal of hematopoietic stem cells. Blood 2006; 108 (13): 4246–54. doi: 10.1182/blood-2006-02-005611.
  39. Singbrant S, Wall M, Moody J, et al. The SKI proto-oncogene enhances the in vivo repopulation of hematopoietic stem cells and causes myeloproliferative disease. Haematologica 2014; 99 (4): 647–55. doi: 10.3324/haematol.2013.093971.
  40. Jeon BN, Yoo JY, Choi WI, et al. Proto-oncogene FBI-1 (Pokemon/ZBTB7A) represses transcription of the tumor suppressor Rb gene via binding competition with Sp1 and recruitment of co-repressors. The Journal of biological chemistry 2008; 283 (48): 33199–210. doi: 10.1074/jbc.M802935200.
  41. Lee SU, Maeda M, Ishikawa Y, et al. LRF-mediated Dll4 repression in erythroblasts is necessary for hematopoietic stem cell maintenance. Blood 2013; 121(6): 918–29.
  42. Yan XQ, Sarmiento U, Sun Y, et al. A novel Notch ligand, Dll4, induces T-cell leukemia/lymphoma when overexpressed in mice by retroviral-mediated gene transfer. Blood 2001; 98 (13): 3793–9. doi: 10.1182/blood.v98.13.3793.
  43. Li J, Tsai MD. Novel insights into the INK4-CDK4/6-Rb pathway: counter action of gankyrin against INK4 proteins regulates the CDK4-mediated phosphorylation of Rb. Biochemistry 2002; 41 (12): 3977–83. doi: 10.1021/bi011550s.
  44. Mine H, Sakurai T, Kashida H, et al. Association of gankyrin and stemness factor expression in human colorectal cancer. Digestive diseases and sciences 2013; 58 (8): 2337–44. doi: 10.1007/s10620-013-2627-8.
  45. Galderisi U, Cipollaro M, Giordano A. The retinoblastoma gene is involved in multiple aspects of stem cell biology. Oncogene 2006; 25 (38): 5250–6. doi: 10.1038/sj.onc.1209736.
  46. Chen PL, Riley DJ, Chen-Kiang S, et al. Retinoblastoma protein directly interacts with and activates the transcription factor NF-IL6. Proc Natl Acad Sci U S A 1996; 93 (1): 465–9. doi: 10.1073/pnas.93.1.465.
  47. Classon M, Kennedy BK, Mulloy R, et al. Opposing roles of pRB and p107 in adipocyte differentiation. Proc Natl Acad Sci U S A 2000; 97 (20): 10826–31. doi: 10.1073/pnas.190343597.
  48. Bergh G, Ehinger M, Olsson I, et al. Involvement of the retinoblastoma protein in monocytic and neutrophilic lineage commitment of human bone marrow progenitor cells. Blood 1999; 94 (6): 1971–8.
  49. Gu W, Schneider JW, Condorelli G, et al. Interaction of myogenic factors and the retinoblastoma protein mediates muscle cell commitment and differentiation. Cell 1993; 72 (3): 309–24. doi: 10.1016/0092-8674(93)90110-c.
  50. Novitch BG, Mulligan GJ, Jacks T, et al. Skeletal muscle cells lacking the retinoblastoma protein display defects in muscle gene expression and accumulate in S and G2 phases of the cell cycle. The Journal of cell biology 1996; 135(2): 441–56. doi: 10.1083/jcb.135.2.441.
  51. Ashe M, Pabon-Pena L, Dees E, et al. LEK1 is a potential inhibitor of pocket protein-mediated cellular processes. The Journal of biological chemistry 2004; 279 (1): 664–76. doi: 10.1074/jbc.M308810200.
  52. Lavender P, Vandel L, Bannister AJ, et al. The HMG-box transcription factor HBP1 is targeted by the pocket proteins and E1A. Oncogene 1997; 14 (22): 2721–8. doi: 10.1038/sj.onc.1201243.
  53. Watanabe N, Kageyama R, Ohtsuka T. Hbp1 regulates the timing of neuronal differentiation during cortical development by controlling cell cycle progression. Development 2015; 142 (13): 2278–90. doi: 10.1242/dev.120477.
  54. Balasenthil S, Vadlamudi RK. Functional interactions between the estrogen receptor coactivator PELP1/MNAR and retinoblastoma protein. The Journal of biological chemistry 2003; 278 (24): 22119–27. doi: 10.1074/jbc.M212822200.
  55. Wang J, Qiang Z, Song S, et al. Temporal expression of Pelp1 during proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells. PLoS One 2013; 8 (10): e75477. doi: 10.1371/journal.pone.0075477.
  56. Wang C, Chen L, Hou X, et al. Interactions between E2F1 and SirT1 regulate apoptotic response to DNA damage. Nature cell biology 2006; 8 (9): 1025–31. doi: 10.1038/ncb1468.
  57. Ryall JG, Dell’Orso S, Derfoul A, The NAD(+)-dependent SIRT1 deacetylase translates a metabolic switch into regulatory epigenetics in skeletal muscle stem cells. Cell Stem Cell 2015; 16 (2): 171–83. doi: 10.1016/j.stem.2014.12.004.
  58. Pradhan S, Kim GD. The retinoblastoma gene product interacts with maintenance human DNA (cytosine-5) methyltransferase and modulates its activity. The EMBO journal 2002; 21 (4): 779–88. doi: 10.1093/emboj/21.4.779.
  59. Trowbridge JJ, Snow JW, Kim J, et al. DNA methyltransferase 1 is essential for and uniquely regulates hematopoietic stem and progenitor cells. Cell Stem Cell 2009; 5 (4): 442–9. doi: 10.1016/j.stem.2009.08.016.
  60. Kim YW, Otterson GA, Kratzke RA, et al. Differential specificity for binding of retinoblastoma binding protein 2 to RB, p107, and TATA-binding protein. Molecular and cellular biology. 1994;14(11):7256–64. doi: 10.1128/mcb.14.11.7256-7264.1994.
  61. Klose RJ, Yan Q, Tothova Z, et al. The Retinoblastoma Binding Protein RBP2 Is an H3K4 Demethylase. Cell 128 (5): 889–900. doi: 10.1016/j.cell.2007.02.013.
  62. Benevolenskaya EV, Murray HL, Branton P, et al. Binding of pRB to the PHD protein RBP2 promotes cellular differentiation. Molecular cell 2005; 18 (6): 623–35. doi: 10.1016/j.molcel.2005.05.012.
  63. Mori T, Ikeda DD, Fukushima T, et al. NIRF constitutes a nodal point in the cell cycle network and is a candidate tumor suppressor. Cell cycle 2011; 10 (19): 3284–99. doi: 10.4161/cc.10.19.17176.
  64. Brehm A, Miska EA, McCance DJ. Retinoblastoma protein recruits histone deacetylase to repress transcription. Nature 1998; 391 (6667): 597–601. doi: 10.1038/35404.
  65. Kretsovali A, Hadjimichael C, Charmpilas N. Histone deacetylase inhibitors in cell pluripotency, differentiation, and reprogramming. Stem Cells Int 2012; 184154 (10): 8. doi: 10.1155/2012/184154.
  66. Endoh M, Endo TA, Endoh T, et al. Polycomb group proteins Ring1A/B are functionally linked to the core transcriptional regulatory circuitry to maintain ES cell identity. Development 2008; 135(8): 1513–24. doi: 10.1242/dev.014340.
  67. Voncken JW, Roelen BA, Roefs M, et al. Rnf2 (Ring1b) deficiency causes gastrulation arrest and cell cycle inhibition. Proc Natl Acad Sci U S A 2003; 100 (5): 2468–73. doi: 10.1073/pnas.0434312100.
  68. Van der Stoop P, Boutsma EA, Hulsman D, et al. Ubiquitin E3 ligase Ring1b/Rnf2 of polycomb repressive complex 1 contributes to stable maintenance of mouse embryonic stem cells. PLoS One 2008; 3 (5): e2235. doi: 10.1371/journal.pone.0002235.
  69. Wang S, Nath N, Adlam M, et al. Prohibitin, a potential tumor suppressor, interacts with RB and regulates E2F function. Oncogene 1999; 18 (23): 3501–10. doi: 10.1038/sj.onc.1202684.
  70. Kowno M, Watanabe-Susaki K, Ishimine H, et al. Prohibitin 2 regulates the proliferation and lineage-specific differentiation of mouse embryonic stem cells in mitochondria. PLoS One 2014; 9 (4): e81552. doi: 10.1371/journal.pone.0081552.
  71. Dunaief JL, Strober BE, Guha S, et al. The retinoblastoma protein and BRG1 form a complex and cooperate to induce cell cycle arrest. Cell 1994; 79(1): 119–30. doi: 10.1016/0092-8674(94)90405-7.
  72. Corsini NS, Sancho-Martinez I, Laudenklos S, et al. The death receptor CD95 activates adult neural stem cells for working memory formation and brain repair. Cell Stem Cell 2009; 5 (2): 178–90. doi: 10.1016/j.stem.2009.05.004.
  73. Holik AZ, Krzystyniak J, Young M, et al. Brg1 is required for stem cell maintenance in the murine intestinal epithelium in a tissue-specific manner. Stem Cells 2013; 31 (11): 2457–66. doi: 10.1002/stem.1498.
  74. Darekar SD, Mushtaq M, Gurrapu S, et al. Mitochondrial ribosomal protein S18-2 evokes chromosomal instability and transforms primary rat skin fibroblasts. Oncotarget 2015; 6 (25): 21016–28. doi: 10.18632/oncotarget.4123.
  75. Nascimento EM, Cox CL, MacArthur S, et al. The opposing transcriptional functions of Sin3A and c-Myc are required to maintain tissue homeostasis. Nature cell biology 2011; 13 (12): 1395–405. doi: 10.1038/ncb2385.
  76. Rao G, Alland L, Guida P, et al. Mouse Sin3A interacts with and can functionally substitute for the amino-terminal repression of the Myc antagonist Mxi1. Oncogene 1996; 12 (5): 1165–72.
  77. Heideman MR, Lancini C, Proost N, et al. Sin3a-associated Hdac1 and Hdac2 are essential for hematopoietic stem cell homeostasis and contribute differentially to hematopoiesis. Haematologica 2014; 99 (8): 1292–303. doi: 10.3324/haematol.2013.092643.
  78. Chen CF, Chen Y, Dai K, et al. A new member of the hsp90 family of molecular chaperones interacts with the retinoblastoma protein during mitosis and after heat shock. Molecular and cellular biology 1996; 16 (9): 4691-9. doi: 10.1128/MCB.16.9.4691.
  79. Wang Y, Lin J, Chen QZ, et al. Overexpression of mitochondrial Hsp75 protects neural stem cells against microglia-derived soluble factor-induced neurotoxicity by regulating mitochondrial permeability transition pore opening in vitro. Int J Mol Med 2015; 36 (6): 1487-96. doi: 10.3892/ijmm.2015.2380.

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