Journal Article
. 2015 May;9(8).
doi: 10.1016/j.molonc.2015.04.008.

Concomitant inactivation of the p53- and pRB- functional pathways predicts resistance to DNA damaging drugs in breast cancer in vivo

Stian Knappskog 1 Elisabet O Berge 2 Ranjan Chrisanthar 2 Stephanie Geisler 2 Vidar Staalesen 2 Beryl Leirvaag 2 Synnøve Yndestad 2 Elise de Faveri 2 Bård O Karlsen 3 David C Wedge 4 Lars A Akslen 5 Peer K Lilleng 6 Erik Løkkevik 7 Steinar Lundgren 8 Bjørn Østenstad 9 Terje Risberg 10 Ingvild Mjaaland 11 Turid Aas 12 Per E Lønning 2 
Affiliations
  • PMID: 26004085
  •     49 References
  •     13 citations

Abstract

Chemoresistance is the main obstacle to cancer cure. Contrasting studies focusing on single gene mutations, we hypothesize chemoresistance to be due to inactivation of key pathways affecting cellular mechanisms such as apoptosis, senescence, or DNA repair. In support of this hypothesis, we have previously shown inactivation of either TP53 or its key activators CHK2 and ATM to predict resistance to DNA damaging drugs in breast cancer better than TP53 mutations alone. Further, we hypothesized that redundant pathway(s) may compensate for loss of p53-pathway signaling and that these are inactivated as well in resistant tumour cells. Here, we assessed genetic alterations of the retinoblastoma gene (RB1) and its key regulators: Cyclin D and E as well as their inhibitors p16 and p27. In an exploratory cohort of 69 patients selected from two prospective studies treated with either doxorubicin monotherapy or 5-FU and mitomycin for locally advanced breast cancers, we found defects in the pRB-pathway to be associated with therapy resistance (p-values ranging from 0.001 to 0.094, depending on the cut-off value applied to p27 expression levels). Although statistically weaker, we observed confirmatory associations in a validation cohort from another prospective study (n = 107 patients treated with neoadjuvant epirubicin monotherapy; p-values ranging from 7.0 × 10(-4) to 0.001 in the combined data sets). Importantly, inactivation of the p53-and the pRB-pathways in concert predicted resistance to therapy more strongly than each of the two pathways assessed individually (exploratory cohort: p-values ranging from 3.9 × 10(-6) to 7.5 × 10(-3) depending on cut-off values applied to ATM and p27 mRNA expression levels). Again, similar findings were confirmed in the validation cohort, with p-values ranging from 6.0 × 10(-7) to 6.5 × 10(-5) in the combined data sets. Our findings strongly indicate that concomitant inactivation of the p53- and pRB- pathways predict resistance towards anthracyclines and mitomycin in breast cancer in vivo.

Keywords: Breast cancer; Resistance; p53; pRB.

Low expression levels of ATM may substitute for CHEK2 /TP53 mutations predicting resistance towards anthracycline and mitomycin chemotherapy in breast cancer.
Stian Knappskog, Ranjan Chrisanthar, +8 authors, Per E Lønning.
Breast Cancer Res, 2012 Mar 17; 14(2). PMID: 22420423    Free PMC article.
Gene expression profiles do not consistently predict the clinical treatment response in locally advanced breast cancer.
Therese Sørlie, Charles M Perou, +8 authors, Per Eystein Lønning.
Mol Cancer Ther, 2006 Nov 24; 5(11). PMID: 17121939
Breast cancer prognostication and prediction in the postgenomic era.
P E Lønning, S Knappskog, +2 authors, J R Lillehaug.
Ann Oncol, 2007 Feb 24; 18(8). PMID: 17317675
Review.
Frequent disruption of the RB pathway in indolent follicular lymphoma suggests a new combination therapy.
Elisa Oricchio, Giovanni Ciriello, +14 authors, Hans-Guido Wendel.
J Exp Med, 2014 Jun 11; 211(7). PMID: 24913233    Free PMC article.
Genetic and epigenetic changes in p21 and p21B do not correlate with resistance to doxorubicin or mitomycin and 5-fluorouracil in locally advanced breast cancer.
Vidar Staalesen, Beryl Leirvaag, Johan Richard Lillehaug, Per Eystein Lønning.
Clin Cancer Res, 2004 May 27; 10(10). PMID: 15161699
TP53 gene mutations predict the response to neoadjuvant treatment with 5-fluorouracil and mitomycin in locally advanced breast cancer.
Stephanie Geisler, Anne-Lise Børresen-Dale, +5 authors, Per Eystein Lønning.
Clin Cancer Res, 2003 Dec 05; 9(15). PMID: 14654539
Topoisomerase IIalpha gene amplification predicts favorable treatment response to tailored and dose-escalated anthracycline-based adjuvant chemotherapy in HER-2/neu-amplified breast cancer: Scandinavian Breast Group Trial 9401.
Scandinavian Breast Group Trial 9401, Minna Tanner, +7 authors, Jonas Bergh.
J Clin Oncol, 2006 May 10; 24(16). PMID: 16682728
Diversity within the pRb pathway: is there a code of conduct?
S Munro, S M Carr, N B La Thangue.
Oncogene, 2012 Jan 18; 31(40). PMID: 22249267
Review.
Concomitant inactivation of the p53- and pRB- functional pathways predicts resistance to DNA damaging drugs in breast cancer in vivo.
Stian Knappskog, Elisabet O Berge, +16 authors, Per E Lønning.
Mol Oncol, 2015 May 26; 9(8). PMID: 26004085    Free PMC article.
Cyclophosphamide dose intensification may circumvent anthracycline resistance of p53 mutant breast cancers.
Jacqueline Lehmann-Che, Fabrice André, +12 authors, Hugues de Thé.
Oncologist, 2010 Mar 17; 15(3). PMID: 20228131    Free PMC article.
Phosphorylation of Ser-20 mediates stabilization of human p53 in response to DNA damage.
N H Chehab, A Malikzay, E S Stavridi, T D Halazonetis.
Proc Natl Acad Sci U S A, 1999 Nov 26; 96(24). PMID: 10570149    Free PMC article.
Highly Cited.
Immortalization of oral keratinocytes by functional inactivation of the p53 and pRb pathways.
Serge J Smeets, Marlon van der Plas, +5 authors, Ruud H Brakenhoff.
Int J Cancer, 2010 May 26; 128(7). PMID: 20499310
Influence of TP53 gene alterations and c-erbB-2 expression on the response to treatment with doxorubicin in locally advanced breast cancer.
S Geisler, P E Lønning, +6 authors, A L Børresen-Dale.
Cancer Res, 2001 Apr 06; 61(6). PMID: 11289122
retrospective analysis of topoisomerase IIa amplifications and deletions as predictive markers in primary breast cancer patients randomly assigned to cyclophosphamide, methotrexate, and fluorouracil or cyclophosphamide, epirubicin, and fluorouracil: Danish Breast Cancer Cooperative Group.
Ann S Knoop, Helle Knudsen, +9 authors, Danish Breast Cancer Cooperative Group.
J Clin Oncol, 2005 Oct 20; 23(30). PMID: 16234514
Activation of ATM and Chk2 kinases in relation to the amount of DNA strand breaks.
Giacomo Buscemi, Paola Perego, +5 authors, Domenico Delia.
Oncogene, 2004 Sep 14; 23(46). PMID: 15361830
Comparisons between different polychemotherapy regimens for early breast cancer: meta-analyses of long-term outcome among 100,000 women in 123 randomised trials.
Early Breast Cancer Trialists' Collaborative Group (EBCTCG), R Peto, +15 authors, K Pritchard.
Lancet, 2011 Dec 14; 379(9814). PMID: 22152853    Free PMC article.
Highly Cited.
Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy.
Hannah Farmer, Nuala McCabe, +11 authors, Alan Ashworth.
Nature, 2005 Apr 15; 434(7035). PMID: 15829967
Highly Cited.
Cyclins and breast cancer.
Robert L Sutherland, Elizabeth A Musgrove.
J Mammary Gland Biol Neoplasia, 2004 Apr 15; 9(1). PMID: 15082921
Review.
Breast cancer subtypes and response to docetaxel in node-positive breast cancer: use of an immunohistochemical definition in the BCIRG 001 trial.
Judith Hugh, John Hanson, +11 authors, Charles Vogel.
J Clin Oncol, 2009 Feb 11; 27(8). PMID: 19204205    Free PMC article.
Highly Cited.
TP53 mutation and p53 overexpression for prediction of response to neoadjuvant treatment in breast cancer patients.
D Kandioler-Eckersberger, C Ludwig, +8 authors, R Jakesz.
Clin Cancer Res, 2000 Feb 03; 6(1). PMID: 10656431
Gene expression and benefit of chemotherapy in women with node-negative, estrogen receptor-positive breast cancer.
Soonmyung Paik, Gong Tang, +11 authors, Norman Wolmark.
J Clin Oncol, 2006 May 25; 24(23). PMID: 16720680
Highly Cited.
Multicenter randomized trial comparing sequential with concomitant administration of doxorubicin and docetaxel as first-line treatment of metastatic breast cancer: a Spanish Breast Cancer Research Group (GEICAM-9903) phase III study.
Emilio Alba, Miguel Martín, +9 authors, Spanish Breast Cancer Research Group.
J Clin Oncol, 2004 Jul 01; 22(13). PMID: 15226326
RB signaling prevents replication-dependent DNA double-strand breaks following genotoxic insult.
Emily E Bosco, Christopher N Mayhew, +3 authors, Erik S Knudsen.
Nucleic Acids Res, 2004 Jan 06; 32(1). PMID: 14704340    Free PMC article.
Study of suboptimum treatment response: lessons from breast cancer.
Per Eystein Lønning.
Lancet Oncol, 2003 Mar 08; 4(3). PMID: 12623363
Review.
Molecular portraits of human breast tumours.
C M Perou, T Sørlie, +15 authors, D Botstein.
Nature, 2000 Aug 30; 406(6797). PMID: 10963602
Highly Cited.
The 70-gene signature as a response predictor for neoadjuvant chemotherapy in breast cancer.
Marieke E Straver, Annuska M Glas, +7 authors, Sjoerd Rodenhuis.
Breast Cancer Res Treat, 2009 Feb 14; 119(3). PMID: 19214742
Microarray analysis reveals a major direct role of DNA copy number alteration in the transcriptional program of human breast tumors.
Jonathan R Pollack, Therese Sørlie, +7 authors, Patrick O Brown.
Proc Natl Acad Sci U S A, 2002 Sep 26; 99(20). PMID: 12297621    Free PMC article.
Highly Cited.
DNA damage-induced activation of p53 by the checkpoint kinase Chk2.
A Hirao, Y Y Kong, +6 authors, T W Mak.
Science, 2000 Mar 10; 287(5459). PMID: 10710310
Highly Cited.
Mapping genetic alterations causing chemoresistance in cancer: identifying the roads by tracking the drivers.
P E Lønning, S Knappskog.
Oncogene, 2013 Mar 12; 32(46). PMID: 23474753
Review.
Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications.
T Sørlie, C M Perou, +14 authors, A L Børresen-Dale.
Proc Natl Acad Sci U S A, 2001 Sep 13; 98(19). PMID: 11553815    Free PMC article.
Highly Cited.
The cyclin D1 high and cyclin E high subgroups of breast cancer: separate pathways in tumorogenesis based on pattern of genetic aberrations and inactivation of the pRb node.
Martin Lodén, Maria Stighall, +4 authors, Göran Landberg.
Oncogene, 2002 Jul 04; 21(30). PMID: 12096344
Phosphorylation of pRB at Ser612 by Chk1/2 leads to a complex between pRB and E2F-1 after DNA damage.
Yasumichi Inoue, Masatoshi Kitagawa, Yoichi Taya.
EMBO J, 2007 Mar 24; 26(8). PMID: 17380128    Free PMC article.
New guidelines to evaluate the response to treatment in solid tumors. European Organization for Research and Treatment of Cancer, National Cancer Institute of the United States, National Cancer Institute of Canada.
P Therasse, S G Arbuck, +8 authors, S G Gwyther.
J Natl Cancer Inst, 2000 Feb 03; 92(3). PMID: 10655437
Highly Cited.
Assessment of response to therapy in advanced breast cancer.
J L Hayward, P P Carbone, +3 authors, R D Rubens.
Br J Cancer, 1977 Mar 01; 35(3). PMID: 856236    Free PMC article.
Highly Cited.
A senescence program controlled by p53 and p16INK4a contributes to the outcome of cancer therapy.
Clemens A Schmitt, Jordan S Fridman, +4 authors, Scott W Lowe.
Cell, 2002 May 23; 109(3). PMID: 12015983
Highly Cited.
Identification and characterization of retinoblastoma gene mutations disturbing apoptosis in human breast cancers.
Elisabet Ognedal Berge, Stian Knappskog, +6 authors, Per Eystein Lønning.
Mol Cancer, 2010 Jul 03; 9. PMID: 20594292    Free PMC article.
A novel type of deletion in the CDKN2A gene identified in a melanoma-prone family.
Stian Knappskog, Jürgen Geisler, +2 authors, Per E Lønning.
Genes Chromosomes Cancer, 2006 Sep 27; 45(12). PMID: 17001621
CHEK2 mutations affecting kinase activity together with mutations in TP53 indicate a functional pathway associated with resistance to epirubicin in primary breast cancer.
Ranjan Chrisanthar, Stian Knappskog, +10 authors, Per Eystein Lønning.
PLoS One, 2008 Aug 30; 3(8). PMID: 18725978    Free PMC article.
Prognostic and predictive value of the 21-gene recurrence score assay in postmenopausal women with node-positive, oestrogen-receptor-positive breast cancer on chemotherapy: a retrospective analysis of a randomised trial.
Kathy S Albain, William E Barlow, +20 authors, Breast Cancer Intergroup of North America.
Lancet Oncol, 2009 Dec 17; 11(1). PMID: 20005174    Free PMC article.
Highly Cited.
Long-term follow-up of patients with complete remission following combination chemotherapy for metastatic breast cancer.
P A Greenberg, G N Hortobagyi, +3 authors, A U Buzdar.
J Clin Oncol, 1996 Aug 01; 14(8). PMID: 8708708
Highly Cited.
Nomograms to predict pathologic complete response and metastasis-free survival after preoperative chemotherapy for breast cancer.
Roman Rouzier, Lajos Pusztai, +13 authors, Gabriel N Hortobagyi.
J Clin Oncol, 2005 Nov 19; 23(33). PMID: 16293864
pRB plays an essential role in cell cycle arrest induced by DNA damage.
E A Harrington, J L Bruce, E Harlow, N Dyson.
Proc Natl Acad Sci U S A, 1998 Sep 30; 95(20). PMID: 9751770    Free PMC article.
Genes causing inherited cancer as beacons to identify the mechanisms of chemoresistance.
Per Eystein Lønning.
Trends Mol Med, 2004 Apr 23; 10(3). PMID: 15102353
Review.
Mutations and polymorphisms of the p21B transcript in breast cancer.
Stian Knappskog, Ranjan Chrisanthar, +4 authors, Per Eystein Lønning.
Int J Cancer, 2007 Apr 20; 121(4). PMID: 17443496
Cellular responses to DNA damage: cell-cycle checkpoints, apoptosis and the roles of p53 and ATM.
T Enoch, C Norbury.
Trends Biochem Sci, 1995 Oct 01; 20(10). PMID: 8533157
Review.
The impact of cyclin D1 overexpression on the prognosis of ER-positive breast cancers: a meta-analysis.
Xiao-Ling Xu, Shu-Zheng Chen, +5 authors, Wei-Min Mao.
Breast Cancer Res Treat, 2013 May 15; 139(2). PMID: 23670132
Regulating the p53 pathway: in vitro hypotheses, in vivo veritas.
Franck Toledo, Geoffrey M Wahl.
Nat Rev Cancer, 2006 Nov 28; 6(12). PMID: 17128209
Highly Cited. Review.
Alterations of the retinoblastoma gene in metastatic breast cancer.
Elisabet Ognedal Berge, Stian Knappskog, Johan Richard Lillehaug, Per Eystein Lønning.
Clin Exp Metastasis, 2011 Jan 22; 28(3). PMID: 21253829    Free PMC article.
Exquisite sensitivity of TP53 mutant and basal breast cancers to a dose-dense epirubicin-cyclophosphamide regimen.
Philippe Bertheau, Elisabeth Turpin, +13 authors, Hugues de Thé.
PLoS Med, 2007 Mar 29; 4(3). PMID: 17388661    Free PMC article.
11a-N-Tosyl-5-deoxi-pterocarpan, LQB-223, a novel compound with potent antineoplastic activity toward breast cancer cells with different phenotypes.
Lauana Greicy Tonon Lemos, Gabriela Nestal de Moraes, +6 authors, Raquel Ciuvalschi Maia.
J Cancer Res Clin Oncol, 2016 Aug 16; 142(10). PMID: 27520309
Prevalence of the CHEK2 R95* germline mutation.
Stian Knappskog, Beryl Leirvaag, +4 authors, Per E Lønning.
Hered Cancer Clin Pract, 2016 Oct 07; 14. PMID: 27708748    Free PMC article.
Heterogeneous perivascular cell coverage affects breast cancer metastasis and response to chemotherapy.
Jiha Kim, Pedro Correa de Sampaio, +11 authors, Valerie S LeBleu.
JCI Insight, 2016 Dec 27; 1(21). PMID: 28018977    Free PMC article.
Dual function of MDM2 and MDMX toward the tumor suppressors p53 and RB.
Jesús Hernández-Monge, Adriana Berenice Rousset-Roman, Ixaura Medina-Medina, Vanesa Olivares-Illana.
Genes Cancer, 2017 Jan 05; 7(9-10). PMID: 28050229    Free PMC article.
Review.
Effects of concomitant inactivation of p53 and pRb on response to doxorubicin treatment in breast cancer cell lines.
Johanna Huun, Per Eystein Lønning, Stian Knappskog.
Cell Death Discov, 2017 Jun 06; 3. PMID: 28580174    Free PMC article.
Protein expression patterns of cell cycle regulators in operable breast cancer.
Flora Zagouri, Vassiliki Kotoula, +21 authors, George Fountzilas.
PLoS One, 2017 Aug 11; 12(8). PMID: 28797035    Free PMC article.
Molecular targeting of the Aurora-A/SMAD5 oncogenic axis restores chemosensitivity in human breast cancer cells.
Mateusz Opyrchal, Malgorzata Gil, +11 authors, Antonino B D'Assoro.
Oncotarget, 2017 Dec 07; 8(53). PMID: 29207686    Free PMC article.
Chemoresistance‑related long non‑coding RNA expression profiles in human breast cancer cells.
Lei Huang, Lihua Zeng, +10 authors, Shui Wang.
Mol Med Rep, 2018 May 12; 18(1). PMID: 29749447    Free PMC article.
Concomitant inactivation of the p53- and pRB- functional pathways predicts resistance to DNA damaging drugs in breast cancer in vivo.
Stian Knappskog, Elisabet O Berge, +16 authors, Per E Lønning.
Mol Oncol, 2015 May 26; 9(8). PMID: 26004085    Free PMC article.
Intrinsic adriamycin resistance in p53-mutated breast cancer is related to the miR-30c/FANCF/REV1-mediated DNA damage response.
Shu Lin, Lifeng Yu, +9 authors, Minjie Wei.
Cell Death Dis, 2019 Sep 13; 10(9). PMID: 31511498    Free PMC article.
Differences in prognosis by p53 expression after neoadjuvant chemotherapy in triple-negative breast cancer.
Soo Youn Bae, Jeong Hyeon Lee, Jeoung Won Bae, Seung Pil Jung.
Ann Surg Treat Res, 2020 Jun 13; 98(6). PMID: 32528908    Free PMC article.
Inflammatory Breast Cancer: Clinical Implications of Genomic Alterations and Mutational Profiling.
Flávia L C Faldoni, Rolando A R Villacis, +10 authors, Silvia R Rogatto.
Cancers (Basel), 2020 Oct 04; 12(10). PMID: 33007869    Free PMC article.
TP53 Mutations in Breast and Ovarian Cancer.
Laxmi Silwal-Pandit, Anita Langerød, Anne-Lise Børresen-Dale.
Cold Spring Harb Perspect Med, 2016 Nov 07; 7(1). PMID: 27815305    Free PMC article.
Review.