Do MRPS18-2 and RB proteins cooperate to control cell stemness and differentiation, preventing cancer development?

In childhood tumors, including retinoblastoma, osteosarcoma, and neuroblastoma, the RB-E2F1 pathway is inactivated, as a rule. These tumors arise from precursor cells that fail to undergo the terminal differentiation. Noteworthy, the RB1-encoded protein (RB) does not control the cell cycle in embryo...

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Опубліковано в: :Experimental Oncology
Дата:2017
Автори: Kashuba, E., Mushtaq, M.
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Опубліковано: Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України 2017
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Цитувати:Do MRPS18-2 and RB proteins cooperate to control cell stemness and differentiation, preventing cancer development? / E. Kashuba, M. Mushtaq // Experimental Oncology. — 2017 — Т. 39, № 1. — С. 12-16. — Бібліогр.: 21 назв. — англ.

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Digital Library of Periodicals of National Academy of Sciences of Ukraine
id nasplib_isofts_kiev_ua-123456789-137599
record_format dspace
spelling Kashuba, E.
Mushtaq, M.
2018-06-17T13:51:03Z
2018-06-17T13:51:03Z
2017
Do MRPS18-2 and RB proteins cooperate to control cell stemness and differentiation, preventing cancer development? / E. Kashuba, M. Mushtaq // Experimental Oncology. — 2017 — Т. 39, № 1. — С. 12-16. — Бібліогр.: 21 назв. — англ.
1812-9269
https://nasplib.isofts.kiev.ua/handle/123456789/137599
In childhood tumors, including retinoblastoma, osteosarcoma, and neuroblastoma, the RB-E2F1 pathway is inactivated, as a rule. These tumors arise from precursor cells that fail to undergo the terminal differentiation. Noteworthy, the RB1-encoded protein (RB) does not control the cell cycle in embryonic stem cells. It has not been yet well understood how RB controls cell stemness and differentiation. The question arises why “inactive” RB is required for the survival and stemness of cells? Recently, we have found that overexpression of the RB-binding protein MRPS18-2 (S18-2) in primary fibroblasts leads to their immortalization, which is accompanied by the induction of embryonic stem cell markers and, eventually, malignant transformation. We suggest that cell stemness may be associated with high expression levels of both proteins, RB and S18-2. There must be a strict regulation of the expression levels of S18-2 and RB during embryogenesis. Disturbances in the expression of these proteins would lead to the abnormalities in development. We think that the S18-2 protein, together with the RB, plays a crucial role in the control on cell stemness and differentiation. We hope to uncover the new mechanisms of the cell fate determination. The S18-2 may serve as a new target for anticancer medicines, which will help to improve human health.
We thank Professor George Klein for the fruitful discussions on the proposed hypothesis.
en
Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
Experimental Oncology
Point of view
Do MRPS18-2 and RB proteins cooperate to control cell stemness and differentiation, preventing cancer development?
Article
published earlier
institution Digital Library of Periodicals of National Academy of Sciences of Ukraine
collection DSpace DC
title Do MRPS18-2 and RB proteins cooperate to control cell stemness and differentiation, preventing cancer development?
spellingShingle Do MRPS18-2 and RB proteins cooperate to control cell stemness and differentiation, preventing cancer development?
Kashuba, E.
Mushtaq, M.
Point of view
title_short Do MRPS18-2 and RB proteins cooperate to control cell stemness and differentiation, preventing cancer development?
title_full Do MRPS18-2 and RB proteins cooperate to control cell stemness and differentiation, preventing cancer development?
title_fullStr Do MRPS18-2 and RB proteins cooperate to control cell stemness and differentiation, preventing cancer development?
title_full_unstemmed Do MRPS18-2 and RB proteins cooperate to control cell stemness and differentiation, preventing cancer development?
title_sort do mrps18-2 and rb proteins cooperate to control cell stemness and differentiation, preventing cancer development?
author Kashuba, E.
Mushtaq, M.
author_facet Kashuba, E.
Mushtaq, M.
topic Point of view
topic_facet Point of view
publishDate 2017
language English
container_title Experimental Oncology
publisher Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
format Article
description In childhood tumors, including retinoblastoma, osteosarcoma, and neuroblastoma, the RB-E2F1 pathway is inactivated, as a rule. These tumors arise from precursor cells that fail to undergo the terminal differentiation. Noteworthy, the RB1-encoded protein (RB) does not control the cell cycle in embryonic stem cells. It has not been yet well understood how RB controls cell stemness and differentiation. The question arises why “inactive” RB is required for the survival and stemness of cells? Recently, we have found that overexpression of the RB-binding protein MRPS18-2 (S18-2) in primary fibroblasts leads to their immortalization, which is accompanied by the induction of embryonic stem cell markers and, eventually, malignant transformation. We suggest that cell stemness may be associated with high expression levels of both proteins, RB and S18-2. There must be a strict regulation of the expression levels of S18-2 and RB during embryogenesis. Disturbances in the expression of these proteins would lead to the abnormalities in development. We think that the S18-2 protein, together with the RB, plays a crucial role in the control on cell stemness and differentiation. We hope to uncover the new mechanisms of the cell fate determination. The S18-2 may serve as a new target for anticancer medicines, which will help to improve human health.
issn 1812-9269
url https://nasplib.isofts.kiev.ua/handle/123456789/137599
citation_txt Do MRPS18-2 and RB proteins cooperate to control cell stemness and differentiation, preventing cancer development? / E. Kashuba, M. Mushtaq // Experimental Oncology. — 2017 — Т. 39, № 1. — С. 12-16. — Бібліогр.: 21 назв. — англ.
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last_indexed 2025-11-26T13:18:03Z
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fulltext 12 Experimental Oncology 39, 12–16, 2017 (March) DO MRPS18-2 AND RB PROTEINS COOPERATE TO CONTROL CELL STEMNESS AND DIFFERENTIATION, PREVENTING CANCER DEVELOPMENT? E. Kashuba1, 2, *, M. Mushtaq1 1Department of Microbiology, Tumor and Cell Biology (MTC), Karolinska Institutet, Stockholm S-17177, Sweden 2R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine, Kyiv 03022, Ukraine In childhood tumors, including retinoblastoma, osteosarcoma, and neuroblastoma, the RB-E2F1 pathway is inactivated, as a rule. These tumors arise from precursor cells that fail to undergo the terminal differentiation. Noteworthy, the RB1-encoded protein (RB) does not control the cell cycle in embryonic stem cells. It has not been yet well understood how RB controls cell stemness and dif- ferentiation. The question arises why “inactive” RB is required for the survival and stemness of cells? Recently, we have found that overexpression of the RB-binding protein MRPS18-2 (S18-2) in primary fibroblasts leads to their immortalization, which is ac- companied by the induction of embryonic stem cell markers and, eventually, malignant transformation. We suggest that cell stem- ness may be associated with high expression levels of both proteins, RB and S18-2. There must be a strict regulation of the expres- sion levels of S18-2 and RB during embryogenesis. Disturbances in the expression of these proteins would lead to the abnormalities in development. We think that the S18-2 protein, together with the RB, plays a crucial role in the control on cell stemness and differentiation. We hope to uncover the new mechanisms of the cell fate determination. The S18-2 may serve as a new target for anticancer medicines, which will help to improve human health. Key Words: mitochondrial ribosomal protein MRPS18-2, retinoblastoma protein RB, stem cells, differentiation, cancer develop- ment, childhood tumors. Retinoblastoma is a sporadic or hereditary child- hood tumor that arises in the retina. Retinoblastoma is usually diagnosed at an early age, such as before the age of two years in cases of hereditary tumor. The age-specific window of retinoblastoma growth suggests that tumor formation depends on the pro- liferation of cells transiently found in the retina (de- veloping retina) [1]. The main genetic background for retinoblastoma is inactivation (mostly deletions) of the RB1 gene [2]. The RB1 gene is the first tumor suppressor gene that was cloned and its deletion was connected with tumor growth. The RB1-encoded protein (RB, NP_000312) is a phosphoprotein expressed uniformly in all tis- sues. The RB is a major regulator of the cell cycle, blocking entry into the S-phase by binding to E2F1. The E2F1 (NP_005216) and several other members of E2F family are transcription factors, which transac- tivate genes required for the entry into the S-phase. RB protein binds to E2F1 and inhibits the transcrip- tional activity of the latter (Fig. 1) [3, 4]. In case of the RB phosphorylation, the E2F1 is released from the protein complex and performs its function. Hence, loss of RB1 leads to uncontrolled cell proliferation. Most probably, this explains also retinoblastoma de- velopment. Importantly, even if the patients with retinoblastoma are cured, they often develop osteosarcomas and melanomas later in life. Osteosarcomas, tumors of the skeleton, arise from the bone-forming cells; usually in the long bones and in the age window of 10–15 years. Genetic conditions of osteosarcomas are similar to retinoblastomas — mainly deletions of RB1 gene, i.e. inactivation of the RB-E2F1 pathway [5]. Melanomas develop from the proliferating mela- noblasts. In melanomas the RB-E2F1 pathway is also inactivated, mainly by alterations in the INK locus on chromosome 9 [6, 7]. This locus encodes two proteins, CDKN2A (p16, NP_000068) and CDKN2B (p15, NP_004927); they bind to and inhibit the cyclin-dependent kinases CDK4 (NP_000066) and CDK6 (NP_001138778). The CDKs phosphorylate RB protein. It was shown that promoter regions of the CDKN2A and CDKN2B genes are heavily methylated in melanomas; consequently, levels of encoded proteins drop dramatically. Phosphorylation of RB is not inhibited then, resulting in the S-phase progression (see Fig. 1). Noteworthy, the E2F1-RB pathway is not functional also in one of the most dangerous childhood tumors, neuroblastoma. Neuroblastoma, sometimes called the embryonic tumor, arises from proliferating neuro- blasts. It was shown that the MYCN (NP_001280157) protein is overexpressed in neuroblastomas. MYCN induces transactivation of the ID2 gene. As a result, the highly expressed ID2 (NP_002157) protein competes with the E2F1 for the binding to RB. As was described above, the free E2F1 promotes cell division [8, 9]. Apart from the inactivation of the RB-E2F1 pathway, and, presumably, enhanced proliferation of cancerous cells, what are the other common features of these four tumor types? Actually, all of the above-mentioned tumors arise from the partially differentiated neural crest stem cells. Thus, retinoblastoma in the eye arises due to prolife- ration of retinoblasts (retinal progenitor cells), which Submitted: August 17, 2016. *Correspondence: E-mail: Elena.Kashuba@ki.se Tel.: +46 8 524 8 67 67 Abbreviations used: RB – RB1-encoded protein; S18-2 – RB-binding protein MRPS18-2. Exp Oncol 2017 39, 1, 12–16 POINT OF VIEw Experimental Oncology 39, 12–16, 2017 (March) 13 should otherwise specialize into optical nerve cells. In other words, the terminal differentiation of retino- blasts is inhibited. Fig. 1. The RB protein is a major regulator of the cell cycle. RB blocks entry into the S-phase by binding to E2F1. The cyclin- dependent kinases CDK4 and CDK6 phosphorylate RB, freeing the E2F1 from a protein complex. This results in S-phase progression Moreover, osteosarcoma results from the prolifera- ting osteoblasts. Normally, osteoblasts should undergo terminal differentiation into osteocytes. Melanoma cells are the proliferating undifferentiated melanoblasts (me- lanocyte precursors). Neuroblastoma is formed by pro- liferating neuroblast precursor cells. Upon the normal developmental process, the neuroblast precursor cells are destined to differentiate into neurons or die by apop- tosis. Hence, all these tumors arise from precursor cells that fail to undergo the terminal stage of differentiation (summarized in Fig. 2). What can be a reason for inhibi- tion of differentiation, in addition to inactivation of the RB-E2F1 pathway, supporting cell division? Yet unexplainably, the RB protein in embryonic stem cells is present mainly in hypo- and hyper- phosphorylated forms, i.e. not in protein complex with the E2F1 [10]. Hence, embryonic cell should proliferate upon such conditions. On the other hand, loss of RB1 leads to embryonic lethality [11]. It is still not well understood how the RB is involved in control on cell stemness and differentiation (see [12] for review). A question arises why the “inactive” RB is re- quired for the maintenance of cell stemness and nor- mal development of a set of tissues? We propose one of the possible explanations. Earlier, we have shown that RB binds to the human mitochondrial ribosomal protein MRPS18-2 (NP_054765, S18-2 in the text). This binding prevents RB-E2F1 complex forma- tion, thus promoting S-phase entry (Fig. 3) [13, 14]. Moreover, overexpression of the S18-2 protein in primary rat embryonic fibroblasts leads to their im- mortalization with the induction of embryonic stem cell markers [15, 16]. Terminally differentiated primary rat skin fibroblasts underwent cell transformation upon ectopic expression of the S18-2 protein. The trans- Fig. 2. Tumors arise from precursor cells that fail to undergo terminal differentiation. The migrating neural crest stem cells are shown in black. The migrating cells differentiate upon arrival to their organ of destination: bone (the upper left panel), skin (the lower left panel), retina (the upper right panel), and the sympathetic nervous system (the lower right panel). If the terminal differentiation of precursor cells is blocked, the tumor arises 14 Experimental Oncology 39, 12–16, 2017 (March) formed cells showed increased telomerase activity, cell cycle disturbance, and chromosomal instability [17]. We concluded that the S18-2 is an oncoprotein and might be involved in carcinogenesis. We have shown recently that the S18-2 protein is expressed at high levels in endometrial cancers compared to hyperplasia and normal endometrium, along with the high level of the free E2F1 [18]. Fig. 3. Mitochondrial ribosomal protein S18-2 binds to RB, preventing the formation of RB-E2F1 complex, thus promoting S-phase entry Our findings and analysis of the published mi- croarray data showed the elevated expression of the S18-2 in stem and cancerous cells. Interestingly, levels of the S18-2 are high in the EBV-transformed B-cells and transformed fibroblasts (Fig. 4, indicated by ar- rows). Besides this, heart, adrenal glands, and skeletal muscles showed the high S18-2 levels. The data used for the analyses described in Fig. 4 were obtained from the GTEx Portal (http://www.gtexportal.org/home) on 27/01/2017. I n t e r e s t i n g l y , u p o n t h e a n a l y s i s of the S18-2 expression pattern in CCLE (Cancer cell line encyclopedia) at Broad Institute web- site (https://portals.broadinstitute.org/ccle), we noticed that the S18-2 expression was quite low in cell lines derived from osteosarcomas, neuro- blastomas, and chondrosarcomas. In melanoma cell lines the S18-2 level was high (Fig. 5, indicated by arrows). The data used for the analyses described in Fig. 5 were obtained from the Broad-Novartis CCLE portal (https://portals.broadinstitute.org/ccle) on 27/01/2017. Noteworthy, in melanoma the RB- E2F1 pathway is inactivated not by RB1 loss, i.e. the RB protein is present. Unfortunately, not much data is reported, concer- ning the molecular mechanisms of chondrosar- coma development. Few authors reported both, loss of RB protein due to loss of heterozygosity of RB1 gene [19], and also methylation of the CDKN2A promoter [20, 21]. The discussed above leads to a hypothesis that the RB and S18-2 proteins are involved together in the maintenance of cell stemness. Hence, the stem cells that could be terminally differentiated should express both, RB and S18-2 proteins. The downregulation of levels or inhibition of the func- tion by binding to other proteins of one or both proteins would lead to a failure in the differentiation of stem cells and, eventually, tumorigenesis. There must be a strict control on the expression of the S18-2 and RB: any disturbance in the expression (upregulation or down- regulation) would result in abnormal proliferation and/ or inhibition of differentiation (Fig. 6). It is feasible to test the proposed hypothesis us- ing, for example the RB1 knockout mouse fibroblasts model. Another possibility is to work with a zebrafish model to knock down S18-2. Also, the sub-lines of can- cer cells derived from retinoblastoma, osteosarcoma, Fig. 4. Expression profile of the S18-2 gene (Entrez Gene ID: 28973). The highest levels of the S18-2 were detected in adrenal gland, heart and skeletal muscles, as well as in transformed fibroblasts and EBV-transformed lymphocytes (indicated with arrows). RPKM – read per kilobase per million mapped reads, normalized to the gene length (see detailed description at http://www.gtexportal.org/home) Experimental Oncology 39, 12–16, 2017 (March) 15 melanoma, and neuroblastoma, expressing the exo- genous S18-2 and RB at the high levels could be gene- rated. The differentiation potential of the obtained cells and their tumorigenicity could be tested then. b a c Fig. 6. RB controls cell stemness and differentiation. (a) RB controls cell stemness and differentiation, keeping the bal- ance of these two processes. Mechanism of regulation is still not well understood. (b) Any disturbance in the RB expression would result in abnormal proliferation and/or inhibition of dif- ferentiation. (c) We hypothesize that RB and S18-2 proteins cooperate to maintain cell stemness. Hence, the stem cells could be characterized by the high expression levels of both, RB and S18-2 proteins. If cells loose RB (or S18-2), no termi- nal differentiation could be achieved. This leads to the tumor development then Concluding, we think that the S18-2 protein, toge- ther with the RB, plays a crucial role in the control on cell stemness and differentiation. We hope to uncover the new mechanisms of the cell fate determination. The S18-2 may probably serve as a new target for anticancer medicines, which will help to improve human health. ACkNOwLEDGMENTS We thank Professor George Klein for the fruitful discussions on the proposed hypothesis. REFERENCES 1. Sage J. The retinoblastoma tumor suppressor and stem cell biology. Genes Dev 2012; 26: 1409–20. 2. Mittnacht S. The retinoblastoma protein – from bench to bedside. Eur J Cell Biol 2005; 84: 97–107. 3. DeCaprio JA, Ludlow JW, Lynch D, et al. The product of the retinoblastoma susceptibility gene has properties of a cell cycle regulatory element. Cell 1989; 58: 1085–95. 4. Shirodkar S, Ewen M, DeCaprio JA, et al. The tran- scription factor E2F interacts with the retinoblastoma product and a p107-cyclin A complex in a cell cycle-regulated manner. Cell 1992; 68: 157–66. 5. Thomas DM, Carty SA, Piscopo DM, et al. The retino- blastoma protein acts as a transcriptional coactivator required for osteogenic differentiation. Mol Cell 2001; 8: 303–16. 6. Bennett DC. Human melanocyte senescence and melanoma susceptibility genes. Oncogene 2003; 22: 3063–9. 7. Hayward NK. Genetics of melanoma predisposition. Oncogene 2003; 22: 3053–62. 8. Iavarone A, Garg P, Lasorella A, et al. The helix-loop- helix protein Id-2 enhances cell proliferation and binds to the retinoblastoma protein. Genes Dev 1994; 8: 1270–84. 9. Lasorella A, Boldrini R, Dominici C, et al. Id2 is critical for cellular proliferation and is the oncogenic effector of N-myc in human neuroblastoma. Cancer Res 2002; 62: 301–6. 10. Conklin JF, Baker J, Sage J. The RB fam- ily is required for the self-renewal and survival of hu- man embryonic stem cells. Nat Commun 2012; 3: 1244. doi:10.1038/ncomms2254. 11. Lipinski MM, Jacks T. The retinoblastoma gene family in differentiation and development. Oncogene 1999; 18: 7873–82. 12. Mushtaq M, Viñas Gaza H, Kashuba EV. Role of the RB interacting proteins in stem cell biology. Adv Cancer Res 2016; 131: 133–57. Fig. 5. Expression profile of the S18-2 gene in immortalized cell lines (Entrez Gene ID: 28973). The lowest levels of the S18-2 were detected in osteosarcoma, chondrosarcoma, and neuroblastoma cell lines (indicated with arrows, the right side of Fig. 5). RMA — Robust multichip averaging, relative units of the signal intensity (see detailed description at https://portals.broadinstitute.org/ccle) 16 Experimental Oncology 39, 12–16, 2017 (March) 13. Snopok B, Yurchenko M, Szekely L, et al. SPR-based immunocapture approach to creating an interfacial sensing architecture: Mapping of the MRS18-2 binding site on reti- noblastoma protein. Anal Bioanal Chem 2006; 386: 2063–73. 14. 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: 5489–94. 15. Kashuba E, Pavan Yenamandra S, Deoram Darekar S, 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: 19866–71. 16. Yenamandra SP, Darekar SD, Kashuba V, et al. Stem cell gene expression in MRPS18-2-immortalized rat embryo- nic fibroblasts. Cell Death Dis 2012; 3: e357. 17. Darekar SD, Mushtaq M, Gurrapu S, et al. Mitochon- drial ribosomal protein S18-2 evokes chromosomal instability and transforms primary rat skin fibroblasts. Oncotarget 2015; 6: 21016–28. 18. Mints M, Mushtaq M, Iurchenko N, et al. Mi- tochondrial ribosomal protein S18-2 is highly expressed in endometrial cancers along with free E2F1. Oncotarget 2016; 7: 22150–8. 19. Röpke M, Boltze C, Meyer B, et al. Rb-loss is associ- ated with high malignancy in chondrosarcoma. Oncol Rep 2006; 15: 89–95. 20. Röpke M, Boltze C, Neumann HW, et al. Genetic and epigenetic alterations in tumor progression in a de- differentiated chondrosarcoma. Pathol Res Pract 2003; 199: 437–44. 21. Franchi A, Baroni G, Sardi I, et al. Dedifferentiated peripheral chondrosarcoma: a clinicopathologic, immuno- histochemical, and molecular analysis of four cases. Virchows Arch 2012; 460: 335–42. Copyright © Experimental Oncology, 2017