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Proliferation-dependent expression and phosphorylation of pRB in B cell non-Hodgkin's lymphomas: dependence on RB1 copy number

Abstract

Some studies have suggested that a significant fraction of non-Hodgkin's lymphomas (NHL) do not express pRB protein, possibly due to deletions of RB1. We examined RB1/centromere 17 copy number by fluorescent in situ hybridisation, and pRB expression/phosphorylation by immunohistochemistry (IHC) and immunoblotting (IB) in 66 cases of B cell NHL. Thirteen cases had lost one RB1 copy relative to centromere 17 copy number and total DNA content. Case 458/88 had no RB1 copies. pRB levels were heterogeneous as assessed by IB (0.04–1.12 relative units), but all tumours, except for case 458/88, expressed pRB localised to the nucleus in >75% of the tumour cells by IHC. The fraction of phosphorylated pRB was correlated with pRB expression (r2 = 0.56, P < 0.001). The 14 cases with loss of RB1 had lower pRB expression (median 0.25) than those without (median 0.48, P < 0.001), but a correlation with S phase fraction (r2 = 0.43, P < 0.001; previously published data for tumour-specific S phase and apoptotic fractions) indicated that the variation in pRB expression was due to differences in proliferative activity. Furthermore, the regression lines for pRB expression vs S phase fraction were not different for the cases with or without loss of one RB1 copy (P = 0.5). Cases 154/88 (one RB1 copy) and 258/88 (two RB1 copies), in addition to case 458/88, had low expression of (hypophosphorylated) pRB (0.04, 0.08 and 0.04), despite their high S phase fractions (21%, 17% and 21%). There was no association between pRB expression/RB1 copy number and apoptotic fraction. Neither pRB expression nor loss of RB1 had prognostic value, but cases 154/88, 258/88, and 458/88 had short survival times (5, 3 and 46 months, respectively) compared to the others (median survival: 44 months, P = 0.03). It is suggested that pRB expression and function are normal in 63 of 66 NHL cases, including 12 of 13 lymphomas with loss of one RB1 allele.

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References

  1. Foon KA, Todd RF . Immunologic classification of leukemia and lymphoma Blood 1986 68: 1–31

    CAS  PubMed  Google Scholar 

  2. Rabbitts TH . Chromosomal translocations in human cancer Nature 1994 372: 143–149

    Article  CAS  PubMed  Google Scholar 

  3. Knutsen T . Cytogenetic changes in the progression of Lymphoma Leuk Lymphoma 1998 31: 1–19

    Article  CAS  PubMed  Google Scholar 

  4. Döhner H, Stilgenbauer S, Benner A, Leupolt E, Kröber A, Bullinger L, Döhner K, Bentz M, Lichter P . Genomic aberrations and survival in chronic lymphocytic leukemia N Engl J Med 2000 343: 1910–1917

    PubMed  Google Scholar 

  5. Peterson L, Lindquist LL, Church S, Kay NE . Frequent clonal abnormalities of chromosome band 13q14 in B-cell chronic lymphocytic leukemia: multiple clones, subclones, and nonclonal alterations in 82 Midwestern patients Genes Chromos Cancer 1992 4: 273–280

    Article  CAS  PubMed  Google Scholar 

  6. Wada M, Okamura T, Okada M, Teramura M, Masuda M, Motoji T, Mizoguchi H . Frequent chromosome arm 13q deletion in aggressive non-Hodgkin's lymphoma Leukemia 1999 13: 792–798

    Article  CAS  PubMed  Google Scholar 

  7. Liu Y, Hermanson M, Grander D, Merup M, Wu X, Heyman M, Rasool O, Juliusson G, Gahrton G, Detlofsson R, Nikiforova N, Buys C, Söderhäll S, Yankovsky N, Zabarovsky E, Einhorn S . 13q deletions in lymphoid malignancies Blood 1995 86: 1911–1915

    CAS  PubMed  Google Scholar 

  8. Liu Y, Szekely L, Grander D, Söderhäll S, Juliusson G, Gahrton G, Linder S, Einhorn S . Chronic lymphocytic leukemia cells with allelic deletions at 13q14 commonly have one intact RB-1 gene: evidence for a role of an adjacent locus Proc Natl Acad Sci USA 1993 90: 8697–8701

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Rosenwald A, Ott G, Krumdiek AK, Dreyling MH, Katzenberger T, Kalla J, Roth S, Ott MM, Muller-Hermelink HK . A biological role for deletions in chromosomal band 13q14 in mantle cell and peripheral T-cell lymphomas? Genes Chromos Cancer 1999 26: 210–214

    Article  CAS  PubMed  Google Scholar 

  10. Stokke T, Erikstein BK, Smhammer L eds Boye E, Steen HB . The retinoblastoma gene product is bound in the nucleus in early G1 phase Exp Cell Res 1993 204: 147–155

    Article  CAS  PubMed  Google Scholar 

  11. Szekely L, Pokrovskaja K, Jiang W-Q, Selivanova G, Löwbeer M, Ringertz N, Wiman KG, Klein G . Resting B-cells, EBV-infected B-blasts and established lymphoblastoid cell lines differ in their Rb, p53 and EBNA-5 expression patterns Oncogene 1995 10: 1869–1874

    CAS  PubMed  Google Scholar 

  12. Kempkes B, Spitkovsky D, Jansen-Durr P, Ellwart JW, Kremmer E, Delecluse HJ, Rottenberger C, Bornkamm GW, Hammerschmidt W . B-cell proliferation and induction of early G1-regulating proteins by Epstein–Barr virus mutants conditional for EBNA2 EMBO J 1995 14: 88–96

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Hollyoake M, Sthler A, Farrell P, Gordon J, Sinclair A . The normal cell cycle activation program is exploited during the infection of quiescent B-lymphocytes by Epstein–Barr virus Cancer Res 1995 55: 4784–4787

    CAS  PubMed  Google Scholar 

  14. Lipinski MM, Jacks T . The retinoblastoma gene family in differentiation and development Oncogene 1999 18: 7873–7882

    Article  CAS  PubMed  Google Scholar 

  15. Hsieh J-K, Fredersdorf S, Kouzarides T, Martin K, Lu X . E2F1-induced apoptosis requires DNA binding but not transactivation and is inhibited by the retinoblastoma protein through direct interaction Genes Dev 1997 11: 1840–1852

    Article  CAS  PubMed  Google Scholar 

  16. Phillips AC, Bates S, Ryan KM, Helin K, Vousden KH . Induction of DNA synthesis and apoptosis are separable functions of E2F1 Genes Dev 1997 11: 1853–1863

    Article  CAS  PubMed  Google Scholar 

  17. Sherr CJ . Cancer cell cycles Science 1996 274: 1672–1677

    Article  CAS  PubMed  Google Scholar 

  18. Nobori T, Mlura K, Wu DJ, Lois A, Takabayashi K, Carson DA . Deletions of the cyclin-dependent kinase-4 inhibitor gene in multiple human cancers Nature 1994 368: 753–756

    Article  CAS  PubMed  Google Scholar 

  19. Hangaishi A, Ogawa S, Imamura N, Miyawaki S, Miura Y, Uike N, Shimazaki C, Emi N, Kunihiko T, Hirosawa S, Kamada N, Kobayashi Y, Takemoto Y, Kitani T, Toyama K, Ohtake S, Yazaki Y, Ueda R, Hirai H . Inactivation of multiple tumour suppressor genes involved in negative regulation of the cell cycle. MTS/p16INK4A/CDKN2, MTS2/p15INK4B, p53, and Rb genes in primary lymphoid malignancies Blood 1996 87: 4949–4958

    CAS  PubMed  Google Scholar 

  20. Wang JYJ, Knudsen ES, Welch PJ . The retinoblastoma suppressor protein Adv Cancer Res 1994 64: 25–85

    Article  CAS  PubMed  Google Scholar 

  21. Horowitz JM, Park S-H, Bogenmann E, Cheng J-C, Yandell DW, Kaye FJ, Minna JD, Dryja TP, Weinberg RA . Frequent inactivation of the retinoblastoma anti-oncogene is restricted to a subset of human tumour cells Proc Natl Acad Sci USA 1990 87: 2775–2779

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Martinez JC, Piris MA, Sanchezbeato M, Villuendas R, Orradre JL, Algara P, Sanchezverde L, Martinez P . Retinoblastoma (Rb) gene-product expression in lymphomas – correlation with Ki67 growth fraction J Pathol 1993 169: 405–412

    Article  CAS  PubMed  Google Scholar 

  23. Weide R, Tiemann M, Pflÿger K-H, Köppler H, Parvizl B, Wacker H-H, Kreipe H-H, Havemann K, Parwaresch MR . Altered expression of the retinoblastoma gene product in human high-grade non-Hodgkin's lymphomas Leukemia 1994 8: 97–101

    CAS  PubMed  Google Scholar 

  24. Kornblau SM, Chen N, del Giglio A, O'Brien S, Deisseroth AB . Retinoblastoma protein expression is frequently altered in chronic lymphocytic leukemia Cancer Res 1994 54: 242–246

    CAS  PubMed  Google Scholar 

  25. Møller MB, Kania PW, Ino Y, Gerdes A-M, Nielsen O, Louis DN, Skjødt K . Frequent disruption of the RB-1 pathway in diffuse large B cell lymphoma: prognostic significance of E2F-1 and p16INK4A Leukemia 2000 14: 898–904

    Article  PubMed  Google Scholar 

  26. Kiviniemi M, Sauroja I, Rajamäki A, Punnonen K, Söderström KO, Salminen E . Cell cycle regulators p27 and pRb in lymphomas – correlation with histology and proliferative activity Br J Cancer 2000 83: 1161–1167

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Jares P, Campo E, Pinyol M, Bosch F, Miquel R, Fernandez PL, Sanchez-Beato M, Soler F, Perez-Losada A, Nayach I, Mallofre C, Piris MA, Montserrat E, Cardesa A . Expression of retinoblastoma gene product (pRb) in mantle cell lymphomas Am J Pathol 1996 148: 1591–1600

    CAS  PubMed  PubMed Central  Google Scholar 

  28. Nguyen PL, Zukerberg LR, Benedict WF, Harris NL . Immunohistochemical detection of p53, bcl-2, and retinoblastoma proteins in follicular lymphoma Am J Clin Pathol 1996 105: 538–543

    Article  CAS  PubMed  Google Scholar 

  29. Geradts J, Andriko JW, Abbondanzo SL . Loss of tumour suppressor gene expression in high-grade but not low-grade non-Hodgkin's lymphomas Am J Clin Pathol 1998 109: 669–674

    Article  CAS  PubMed  Google Scholar 

  30. Grierson AJ, Hodgkins MA, Hancock BW, Goepel JR, Royds J, Goyns MH . Investigation of the RB-1 tumour suppressor gene in a United Kingdom series of non-Hodgkin's lymphomas Leuk Lymphoma 1996 23: 353–363

    Article  CAS  PubMed  Google Scholar 

  31. Ballerini P, Gaidano G, Gong JZ, Tassi V, Saglio G, Knowles DM, Dalla-Favera R . Multiple genetic lesions in acquired immunodeficiency syndrome-related non-Hodgkin's lymphoma Blood 1993 81: 166–176

    CAS  PubMed  Google Scholar 

  32. Scheffner M, Munger K, Byrne JC, Howley PM . The state of the p53 and retinoblastoma genes in human cervical carcinoma cell lines Proc Natl Acad Sci USA 1991 88: 5523–5527

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Horowitz JM, Yandell DW, Park S-H, Canning S, Whyte P, Buchkovich K, Harlow E, Weinberg RA, Dryja TP . Point mutational inactivation of the retinoblastoma antioncogene Science 1989 243: 937–940

    Article  CAS  PubMed  Google Scholar 

  34. Mittnacht S, Weinberg RA . G1/S phosphorylation of the retinoblastoma protein is associated with an altered affinity for the nuclear compartment Cell 1991 65: 381–393

    Article  CAS  PubMed  Google Scholar 

  35. Chellappan SP, Hiebert S, Mudryj M, Horowitz JM, Nevins JR . The E2F transcription factor is a cellular target for the RB protein Cell 1991 65: 1053–1061

    Article  CAS  PubMed  Google Scholar 

  36. Dreyling MH, Bullinger L, Ott G, Stilgenbauer S, Müller-Hermelink HK, Bentz M, Hiddelmann W, Döhner H . Alterations of the cyclin D1/p16-pRB pathway in mantle cell lymphoma Cancer Res 1997 57: 4608–4614

    CAS  PubMed  Google Scholar 

  37. Galteland E, Holte H, Stokke T . c-MYC, RB-1, TP53, and centromere 8 and 17 copy number in B-cell non-Hodgkin's lymphomas assessed by dual-color fluorescence in situ hybridisation Cytometry 1999 38: 53–60

    Article  CAS  PubMed  Google Scholar 

  38. Stokke T, Galteland E, Holte H, Smhammer L eds Suo Z, Smeland EB, Børresen-Dale AL, Deangelis P, Steen HB . Oncogenic aberrations in the p53 pathway are associated with a high S phase fraction and poor patient survival in B-cell non-Hodgkin's lymphoma Int J Cancer 2000 89: 313–324

    Article  CAS  PubMed  Google Scholar 

  39. Stokke T, Holte H, Smhammer L eds Smeland EB, Kaalhus O, Steen HB . Proliferation and apoptosis in malignant and normal cells in B-cell non-Hodgkin's lymphomas Br J Cancer 1998 77: 1832–1838

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Holte H, Suo Z, Smeland EB, Kvaløy S, Langholm R, Stokke T . The prognostic value of lymphoma-specific S-phase compared with other cell proliferation markers Acta Oncologica 1999 38: 495–503

    Article  CAS  PubMed  Google Scholar 

  41. DeAngelis P, Stokke T, Smhammer L eds Lothe RA, Lehne G, Chen Y, Clausen OP . P-glycoprotein is not expressed in a majority of colorectal carcinomas and is not regulated by mutant p53 in vivo Br J Cancer 1995 72: 307–311

    Article  CAS  Google Scholar 

  42. Lømo J, Smeland EB, Krajewski S, Reed JC, Blomhoff HK . Expression of the Bcl-2 homologue Mcl-1 correlates with survival of peripheral blood B-lymphocytes Cancer Res 1996 56: 40–43

    PubMed  Google Scholar 

  43. Kraggerud SM, Jacobsen KD, Berner Å, Stokke T, Holm R, Smhammer L eds Børresen-Dale AL, Fosså SD . A comparison of different modes for the detection of p53 accumulation. A study of bladder cancer Path Res Pract 1997 193: 471–478

    Article  CAS  PubMed  Google Scholar 

  44. Stokke T, DeAngelis P, Smhammer L eds Galteland E, Steen HB, Smeland EB, Delabie J, Holte H . Loss of chromosome 11q21-23.1 and 17p and gain of chromosome 6p are independent prognostic indicators in B-cell Non-Hodgkin's lymphoma Br J Cancer 2001 85: 1900–1913

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Cuneo A, Bigoni R, Rigolin GM, Roberti MG, Bardi A, Campioni D, Minotto C, Agostini P, Milani R, Bullrich F, Negrini M, Croce C, Castoldi G . 13q14 deletion in non-Hodgkin's lymphoma: correlation with clinicopathologic features Haematologica 1999 84: 589–593

    CAS  PubMed  Google Scholar 

  46. Lens D, Matutes E, Catovsky D, Coignet LJA . Frequent deletions at 11q23 and 13q14 in B cell prolymphocytic leukemia (B-PLL) Leukemia 2000 14: 427–430

    Article  CAS  PubMed  Google Scholar 

  47. Tanaka K, Arif M, Eguchi M, Guo SX, Hayashi Y, Asaoku H, Kyo T, Dohy H, Kamada N . Frequent allelic loss of the RB, D13S319 and D13S25 locus in myeloid malignancies with deletion/translocation at 13q14 of chromosome 13, but not in lymphoid malignancies Leukemia 1999 13: 1367–1373

    Article  CAS  PubMed  Google Scholar 

  48. Cook WD, McCaw BJ . Accommodating haploinsufficient tumour suppressor genes in Knudsen's model Oncogene 2000 19: 3434–3438

    Article  CAS  PubMed  Google Scholar 

  49. Nakamura M, Sakaki T, Hashimoto H, Nakase H, Ishida E, Shimada K, Konishi N . Frequent alterations of the p14 ARF and p16INK4a genes in primary central nervous system lymphomas Cancer Res 2001 61: 6335–6339

    CAS  PubMed  Google Scholar 

  50. Koduru PRK, Raju K, Vadmal V, Menezes G, Shah S, Susin M, Kolitz J, Broome JD . Correlation between mutation in p53, p53 expression, cytogenetics, histologic type, and survival in patients with B-cell non-Hodgkin's lymphoma Blood 1997 90: 4078–4091

    CAS  PubMed  Google Scholar 

  51. Louie DC, Offit K, Jaslow R, Parsa NZ, Murty VV, Schluger A, Chaganti RS . p53 expression as a marker of poor prognosis in mantle cell lymphomas with t(11;14)(q13;q32) Blood 1995 86: 2892–2899

    CAS  PubMed  Google Scholar 

  52. Møller MB, Ino Y, Gerdes A-M, Skjødt K, Louis DN, Pedersen NT . Aberrations of the p53 pathway components p53, MDM2 and CDKN2A appear independent in diffuse large B cell lymphoma Leukemia 1999 13: 453–459

    Article  PubMed  Google Scholar 

  53. Grønbæk K, de Nully Brown P, Møller MB, Nedergaard T, Ralfkiaer E, Møller P, Zeuthen J, Guldberg P . Concurrent disruption of p16INK4a and the ARF-p53 pathway predicts poor prognosis in aggressive non-Hodgkin's lymphoma Leukemia 2000 14: 1727–1735

    Article  PubMed  Google Scholar 

  54. Sherr CJ, Roberts JM . Inhibitors of mammalian G1 cyclin-dependent kinases Genes Dev 1995 9: 1149–1163

    Article  CAS  PubMed  Google Scholar 

  55. Elledge SJ, Harper JW . Cdk inhibitors: on the threshold of checkpoints and development Curr Opin Cell Biol 1994 6: 847–852

    Article  CAS  PubMed  Google Scholar 

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This study was sponsored by the Norwegian Cancer Society.

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Galteland, E., Smedshammer, L., Suo, Z. et al. Proliferation-dependent expression and phosphorylation of pRB in B cell non-Hodgkin's lymphomas: dependence on RB1 copy number. Leukemia 16, 1549–1555 (2002). https://doi.org/10.1038/sj.leu.2402644

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