Home Transcript expression and genetic variability analysis of caspases in breast carcinomas suggests CASP9 as the most interesting target
Article
Licensed
Unlicensed Requires Authentication

Transcript expression and genetic variability analysis of caspases in breast carcinomas suggests CASP9 as the most interesting target

  • Veronika Brynychova , Viktor Hlavac , Marie Ehrlichova , Radka Vaclavikova , Vlasta Nemcova-Furstova , Vaclav Pecha , Marketa Trnkova , Marcela Mrhalova , Roman Kodet , David Vrana , Jiri Gatek , Marie Bendova , Zdenka Vernerova , Jan Kovar and Pavel Soucek EMAIL logo
Published/Copyright: June 21, 2016
Become an author with De Gruyter Brill
Submit Manuscript Author Information
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 55 Issue 1

Abstract

Background:

Apoptosis plays a critical role in cancer cell survival and tumor development. We provide a hypothesis-generating screen for further research by exploring the expression profile and genetic variability of caspases (2, 3, 7, 8, 9, and 10) in breast carcinoma patients. This study addressed isoform-specific caspase transcript expression and genetic variability in regulatory sequences of caspases 2 and 9.

Methods:

Gene expression profiling was performed by quantitative real-time PCR in tumor and paired non-malignant tissues of two independent groups of patients. Genetic variability was determined by high resolution melting, allelic discrimination, and sequencing analysis in tumor and peripheral blood lymphocyte DNA of the patients.

Results:

CASP3 A+B and S isoforms were over-expressed in tumors of both patient groups. The CASP9 transcript was down-regulated in tumors of both groups of patients and significantly associated with expression of hormonal receptors and with the presence of rs4645978-rs2020903-rs4646034 haplotype in the CASP9 gene. Patients with a low intratumoral CASP9A/B isoform expression ratio (predicted to shift equilibrium towards anti-apoptotic isoform) subsequently treated with adjuvant chemotherapy had a significantly shorter disease-free survival than those with the high ratio (p=0.04). Inheritance of CC genotype of rs2020903 in CASP9 was associated with progesterone receptor expression in tumors (p=0.003).

Conclusions:

Genetic variability in CASP9 and expression of its splicing variants present targets for further study.

Keywords: alternative splicing; breast carcinoma; caspases; prognosis; transcript

  1. Author contributions:All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission. No writing assistance was utilized in the production of this manuscript.

  2. Research funding: This work was supported by grant of the CU Grant Agency no.: 1444313 to V.B., MH CZ – DRO (National Institute of Public Health – NIPH, 75010330) project, and grant of the Internal Grant Agency of the Czech Ministry of Health, no.: 14055-3 to P.S.

  3. Employment or leadership:None declared.

  4. Honorarium: None declared.

  5. Competing interests:The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

References

1. Ferlay J, Soerjomataram I, Ervik M, Dikshit R, Eser S, Mathers C, et al. GLOBOCAN 2012 v1.0, Cancer incidence and mortality worldwide: IARC CancerBase No. 11 [Internet]. Lyon, France: International Agency for Research on Cancer; 2013. Available at: http://globocan.iarc.fr. Accessed: 30 Oct 2015.Search in Google Scholar

2. Murray S, Briasoulis E, Linardou H, Bafaloukos D, Papadimitriou C. Taxane resistance in breast cancer: mechanisms, predictive biomarkers and circumvention strategies. Cancer Treat Rev 2009;38:890–903.10.1016/j.ctrv.2012.02.011Search in Google Scholar PubMed

3. Olsson M, Zhivotovsky B. Caspases and cancer. Cell Death Differ 2011;18:1441–9.10.1038/cdd.2011.30Search in Google Scholar PubMed PubMed Central

4. Labi V, Erlacher M. How cell death shapes cancer. Cell Death Dis 2015;6:e1675.10.1038/cddis.2015.20Search in Google Scholar PubMed PubMed Central

5. Walsh JG, Cullen SP, Sheridan C, Lüthi AU, Gerner C, Martin SJ. Executioner caspase-3 and caspase-7 are functionally distinct proteases. Proc Natl Acad Sci USA 2008;105:12815–9.10.1073/pnas.0707715105Search in Google Scholar PubMed PubMed Central

6. Vakifahmetoglu-Norberg H, Zhivotovsky B. The unpredictable caspase-2: what can it do? Trends Cell Biol 2010;20:150–9.10.1016/j.tcb.2009.12.006Search in Google Scholar PubMed

7. Kovář J, Ehrlichova M, Smejkalova B, Zanardi I, Ojima I, Gut I. Comparison of cell death-inducing effect of novel taxane SB-T-1216 and paclitaxel in breast cancer cells. Anticancer Res 2009;29:2951–60.Search in Google Scholar

8. Mielgo A, Torres VA, Clair K, Barbero S, Stupack DG. Paclitaxel promotes a caspase 8-mediated apoptosis through death effector domain association with microtubules. Oncogene 2009;28:3551–62.10.1038/onc.2009.210Search in Google Scholar PubMed PubMed Central

9. Vobořilová J, Němcová-Fürstová V, Neubauerová J, Ojima I, Zanardi I, Gut I, et al. Cell death induced by novel fluorinated taxanes in drug-sensitive and drug-resistant cancer cells. Invest New Drugs 2011;29:411–23.10.1007/s10637-009-9368-8Search in Google Scholar PubMed PubMed Central

10. Brynychová V, Hlaváč V, Ehrlichová M, Václavíková R, Pecha V, Trnková M, et al. Importance of transcript levels of caspase-2 isoforms S and L for breast carcinoma progression. Future Oncol 2013;9:427–38.10.2217/fon.12.200Search in Google Scholar PubMed

11. Huang Y, Shin NH, Sun Y, Wang KK. Molecular cloning and characterization of a novel caspase-3 variant that attenuates apoptosis induced by proteasome inhibition. Bioch Biophys Res Commun 2001;283:762–9.10.1006/bbrc.2001.4871Search in Google Scholar PubMed

12. Horiuchi T, Himeji D, Tsukamoto H, Harashima SI, Hashimura C, Hayashi K. Dominant expression of a novel splice variant of caspase-8 in human peripheral blood lymphocytes. Bioch Biophys Res Commun 2000;272:877–81.10.1006/bbrc.2000.2841Search in Google Scholar PubMed

13. Himeji D, Horiuchi T, Tsukamoto H, Hayashi K, Watanabe T, Harada M. Characterization of caspase-8L: a novel isoform of caspase-8 that behaves as an inhibitor of the caspase cascade. Blood 2002;99:4070–8.10.1182/blood.V99.11.4070Search in Google Scholar PubMed

14. Seol DW, Billiar TR. A caspase-9 variant missing the catalytic site is an endogenous inhibitor of apoptosis. J Biol Chem 1999;274:2072–6.10.1074/jbc.274.4.2072Search in Google Scholar PubMed

15. Shultz JC, Goehe RW, Murudkar CS, Wijesinghe DS, Mayton EK, Massiello A, et al. SRSF1 regulates the alternative splicing of caspase 9 via a novel intronic splicing enhancer affecting the chemotherapeutic sensitivity of non–small cell lung cancer cells. Mol Cancer Res 2011;9:889–900.10.1158/1541-7786.MCR-11-0061Search in Google Scholar PubMed PubMed Central

16. Ghavami S, Hashemi M, Ande SR, Yeganeh B, Xiao W, Eshraghi M, et al. Apoptosis and cancer: mutations within caspase genes. J Med Genet 2009;46:497–510.10.1136/jmg.2009.066944Search in Google Scholar PubMed

17. Theodoropoulos GE, Michalopoulos NV, Pantou MP, Kontogianni P, Gazouli M, Karantanos T, et al. Caspase 9 promoter polymorphisms confer increased susceptibility to breast cancer. Cancer Genet 2012;205:508–12.10.1016/j.cancergen.2012.08.001Search in Google Scholar PubMed

18. Hubackova M, Vaclavikova R, Ehrlichova M, Mrhalova M, Kodet R, Kubackova K, et al. Association of superoxide dismutases and NAD (P) H quinone oxidoreductases with prognosis of patients with breast carcinomas. Int J Cancer 2012;130:338–48.10.1002/ijc.26006Search in Google Scholar PubMed

19. Tavassoli FA, Devilee P, editors. Pathology and genetics: tumours of the breast and female genital organs, Vol. 4. Lyon: IARC Press, 2003.Search in Google Scholar

20. Goldhirsch A, Winer EP, Coates AS, Gelber RD, Piccart-Gebhart M, Thürlimann B, et al. Personalizing the treatment of women with early breast cancer: highlights of the St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2013. Ann Oncol 2013;24:2206–23.10.1093/annonc/mdt303Search in Google Scholar PubMed PubMed Central

21. Topić E, Gluhak J. Isolation of restrictible DNA. Eur J Clin Chem Clin Biochem 1991;29:327–30.Search in Google Scholar

22. Côté J, Dupuis S, Jiang ZH, Wu JY. Caspase-2 pre-mRNA alternative splicing: identification of an intronic element containing a decoy 3′ acceptor site. Proc Natl Acad Sci 2001;98:938–43.10.1073/pnas.98.3.938Search in Google Scholar PubMed PubMed Central

23. Logette E, Wotawa A, Solier S, Desoche L, Solary E, Corcos L. The human caspase-2 gene: alternative promoters, pre-mRNA splicing and AUG usage direct isoform-specific expression. Oncogene 2003;22:935–46.10.1038/sj.onc.1206172Search in Google Scholar PubMed

24. Hlaváč V, Brynychová V, Václavíková R, Ehrlichová M, Vrána D, Pecha V, et al. The expression profile of ATP-binding cassette transporter genes in breast carcinoma. Pharmacogenomics 2013;14:515–29.10.2217/pgs.13.26Search in Google Scholar PubMed

25. Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, et al. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 2009;55:611–22.10.1373/clinchem.2008.112797Search in Google Scholar PubMed

26. Ward LD, Kellis M. HaploReg: a resource for exploring chromatin states, conservation, and regulatory motif alterations within sets of genetically linked variants. Nucleic Acid Res 2012;40:D930–4.10.1093/nar/gkr917Search in Google Scholar PubMed PubMed Central

27. Hlaváč V, Brynychová V, Václavíková R, Ehrlichová M, Vrána D, Pecha V, et al. The role of cytochromes p450 and aldo-keto reductases in prognosis of breast carcinoma patients. Medicine 2014;93:e255.10.1097/MD.0000000000000255Search in Google Scholar PubMed PubMed Central

28. Benjamini Y, Hochberg Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J Roy Stat Soc B Met 1995;57:289–300.10.1111/j.2517-6161.1995.tb02031.xSearch in Google Scholar

29. Srinivasula SM, Guo AM, Zhan Y, Lazebnik Y, Fernandes-Alnemri T, Alnemri ES. Identification of an endogenous dominant-negative short isoform of caspase-9 that can regulate apoptosis. Cancer Res 1999;59:999–1002.Search in Google Scholar

30. Liu JR, Opipari AW, Tan L, Jiang Y, Zhang Y, Tang H, et al. Dysfunctional apoptosome activation in ovarian cancer implications for chemoresistance. Cancer Res 2002;62:924–31.Search in Google Scholar

31. Lobenhofer EK, Bennett L, Cable LA, Li L, Bushel PR, Afshari CA. Regulation of DNA replication fork genes by 17β-estradiol. Mol Endocrinol 2002;16:1215–29.10.1210/mend.16.6.0858Search in Google Scholar PubMed

32. Frasor J, Danes JM, Komm B, Chang KC, Lyttle CR, Katzenellenbogen BS. Profiling of estrogen up-and down-regulated gene expression in human breast cancer cells: insights into gene networks and pathways underlying estrogenic control of proliferation and cell phenotype. Endocrinology 2003;144:4562–74.10.1210/en.2003-0567Search in Google Scholar PubMed

33. Rodríguez-Berriguete G, Galvis L, Fraile B, de Bethencourt FR, Martínez-Onsurbe P, Olmedilla G, et al. Immunoreactivity to caspase-3, caspase-7, caspase-8, and caspase-9 forms is frequently lost in human prostate tumors. Hum Pathol 2012;43:229–37.10.1016/j.humpath.2011.04.024Search in Google Scholar PubMed

34. Ehrlichova M, Mohelnikova-Duchonova B, Hrdy J, Brynychova V, Mrhalova M, Kodet R, et al. The association of taxane resistance genes with the clinical course of ovarian carcinoma. Genomics 2013;102:96–101.10.1016/j.ygeno.2013.03.005Search in Google Scholar PubMed

35. Xu W, Jiang S, Xu Y, Chen B, Li Y, Zong F, et al. A meta-analysis of caspase 9 polymorphisms in promoter and exon sequence on cancer susceptibility. PLoS One 2012;7:e37443.10.1371/journal.pone.0037443Search in Google Scholar PubMed PubMed Central

36. Kouros-Mehr H, Slorach EM, Sternlicht MD, Werb Z. GATA-3 maintains the differentiation of the luminal cell fate in the mammary gland. Cell 2006;127:1041–55.10.1016/j.cell.2006.09.048Search in Google Scholar PubMed PubMed Central

37. Wang D, Lu P, Zhang H, Luo M, Zhang X, Wei X, et al. Oct-4 and Nanog promote the epithelial-mesenchymal transition of breast cancer stem cells and are associated with poor prognosis in breast cancer patients. Oncotarget 2014;5:10803–15.10.18632/oncotarget.2506Search in Google Scholar PubMed PubMed Central

38. Vakkala M, Pääkkö P, Soini Y. Expression of caspases 3, 6 and 8 is increased in parallel with apoptosis and histological aggressiveness of the breast lesion. Br J Cancer 1999;81:592.10.1038/sj.bjc.6690735Search in Google Scholar PubMed PubMed Central

39. O’Donovan N, Crown J, Stunell H, Hill AD, McDermott E, O’Higgins N, et al. Caspase 3 in breast cancer. Clin Cancer Res 2003;9:738–42.Search in Google Scholar

40. Végran F, Boidot R, Oudin C, Riedinger JM, Bonnetain F, Lizard-Nacol S. Overexpression of caspase-3s splice variant in locally advanced breast carcinoma is associated with poor response to neoadjuvant chemotherapy. Clin Cancer Res 2006;12:5794–800.10.1158/1078-0432.CCR-06-0725Search in Google Scholar PubMed

41. Nassar A, Lawson D, Cotsonis G, Cohen C. Survivin and caspase-3 expression in breast cancer: correlation with prognostic parameters, proliferation, angiogenesis, and outcome. Appl Immunohisto M M 2008;16:113–20.10.1097/PAI.0b013e318032ea73Search in Google Scholar PubMed

42. Vinothini G, Murugan RS, Nagini S. Mitochondria-mediated apoptosis in patients with adenocarcinoma of the breast: correlation with histological grade and menopausal status. Breast 2011;20:86–92.10.1016/j.breast.2010.08.002Search in Google Scholar PubMed

43. Stephens PJ, Tarpey PS, Davies H, Van Loo P, Greenman C, Wedge DC, et al. The landscape of cancer genes and mutational processes in breast cancer. Nature 2012;486:400–4.10.1038/nature11017Search in Google Scholar PubMed PubMed Central

Supplemental Material:

The online version of this article (DOI: 10.1515/cclm-2016-0271) offers supplementary material, available to authorized users.

Received: 2016-4-5
Accepted: 2016-5-17
Published Online: 2016-6-21
Published in Print: 2017-1-1

©2017 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Describing children’s changes using clinical chemistry analytes
  4. Review
  5. Assessment of vitamin D status – a changing landscape
  6. Opinion Papers
  7. Order of blood draw: Opinion Paper by the European Federation for Clinical Chemistry and Laboratory Medicine (EFLM) Working Group for the Preanalytical Phase (WG-PRE)
  8. The principles of Health Technology Assessment in laboratory medicine
  9. Genetics and Molecular Diagnostics
  10. Functional polymorphism of lncRNA MALAT1 contributes to pulmonary arterial hypertension susceptibility in Chinese people
  11. General Clinical Chemistry and Laboratory Medicine
  12. Influence of pneumatic tube system transport on routinely assessed and spectrophotometric cerebrospinal fluid parameters
  13. Estimation of hematocrit in filter paper dried bloodspots by potassium measurement: advantage of use of perimeter ring samples over circular center sub-punch samples
  14. Evaluation of the first fully automated immunoassay method for the measurement of stimulating TSH receptor autoantibodies in Graves’ disease
  15. Comparison of four immunoassays to measure serum ferritin concentrations and iron deficiency prevalence among non-pregnant Cambodian women and Congolese children
  16. Activity and expression of dipeptidyl peptidase IV on peripheral blood mononuclear cells in patients with early steroid and disease modifying antirheumatic drugs naïve rheumatoid arthritis
  17. Hematology and Coagulation
  18. Neutrophil CD64 Index as a superior biomarker for early diagnosis of infection in febrile patients in the hematology department
  19. Dilute Russell’s viper venom time reagents in lupus anticoagulant testing: a well-considered choice
  20. Reference Values and Biological Variations
  21. Pediatric reference intervals for alkaline phosphatase
  22. Cancer Diagnostics
  23. Transcript expression and genetic variability analysis of caspases in breast carcinomas suggests CASP9 as the most interesting target
  24. CDH1 (E-cadherin) expression independently affects clinical outcome in acute myeloid leukemia with normal cytogenetics
  25. Cardiovascular Diseases
  26. Association of monocyte to HDL cholesterol level with contrast induced nephropathy in STEMI patients treated with primary PCI
  27. Low concentrations of adropin are associated with endothelial dysfunction as assessed by flow-mediated dilatation in patients with metabolic syndrome
  28. Preoperative serum potassium predicts the clinical outcome after non-cardiac surgery
  29. Diabetes
  30. A simple colorimetric assay for measuring fructosamine 3 kinase activity
  31. Acknowledgment
  32. Acknowledgment
  33. Letters to the Editor
  34. Normalisation issues in glucose measurements using phlebotomy tubes with liquid additives
  35. Utility of dilution tests in investigating interference in the free thyroxine assay
  36. Plasminogen activator inhibitor-1 as a marker of cardiovascular response in professional mountain ultra-marathon runners
  37. Pitfalls of LSD screening assays: comparison of KIMS and CEDIA immunoassays with LC-MS
  38. A two site comparison of two point-of-care activated clotting time systems
  39. Detection of the common South-East Asian β0-thalassemia mutations in samples with borderline HbA2 levels
  40. Biclonal plasma cell myeloma with the simultaneous appearance of both secretory lambda and nonsecretory kappa monoclonal light chains
https://doi.org/10.1515/cclm-2016-0271
Keywords for this article
alternative splicing; breast carcinoma; caspases; prognosis; transcript

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Describing children’s changes using clinical chemistry analytes
  4. Review
  5. Assessment of vitamin D status – a changing landscape
  6. Opinion Papers
  7. Order of blood draw: Opinion Paper by the European Federation for Clinical Chemistry and Laboratory Medicine (EFLM) Working Group for the Preanalytical Phase (WG-PRE)
  8. The principles of Health Technology Assessment in laboratory medicine
  9. Genetics and Molecular Diagnostics
  10. Functional polymorphism of lncRNA MALAT1 contributes to pulmonary arterial hypertension susceptibility in Chinese people
  11. General Clinical Chemistry and Laboratory Medicine
  12. Influence of pneumatic tube system transport on routinely assessed and spectrophotometric cerebrospinal fluid parameters
  13. Estimation of hematocrit in filter paper dried bloodspots by potassium measurement: advantage of use of perimeter ring samples over circular center sub-punch samples
  14. Evaluation of the first fully automated immunoassay method for the measurement of stimulating TSH receptor autoantibodies in Graves’ disease
  15. Comparison of four immunoassays to measure serum ferritin concentrations and iron deficiency prevalence among non-pregnant Cambodian women and Congolese children
  16. Activity and expression of dipeptidyl peptidase IV on peripheral blood mononuclear cells in patients with early steroid and disease modifying antirheumatic drugs naïve rheumatoid arthritis
  17. Hematology and Coagulation
  18. Neutrophil CD64 Index as a superior biomarker for early diagnosis of infection in febrile patients in the hematology department
  19. Dilute Russell’s viper venom time reagents in lupus anticoagulant testing: a well-considered choice
  20. Reference Values and Biological Variations
  21. Pediatric reference intervals for alkaline phosphatase
  22. Cancer Diagnostics
  23. Transcript expression and genetic variability analysis of caspases in breast carcinomas suggests CASP9 as the most interesting target
  24. CDH1 (E-cadherin) expression independently affects clinical outcome in acute myeloid leukemia with normal cytogenetics
  25. Cardiovascular Diseases
  26. Association of monocyte to HDL cholesterol level with contrast induced nephropathy in STEMI patients treated with primary PCI
  27. Low concentrations of adropin are associated with endothelial dysfunction as assessed by flow-mediated dilatation in patients with metabolic syndrome
  28. Preoperative serum potassium predicts the clinical outcome after non-cardiac surgery
  29. Diabetes
  30. A simple colorimetric assay for measuring fructosamine 3 kinase activity
  31. Acknowledgment
  32. Acknowledgment
  33. Letters to the Editor
  34. Normalisation issues in glucose measurements using phlebotomy tubes with liquid additives
  35. Utility of dilution tests in investigating interference in the free thyroxine assay
  36. Plasminogen activator inhibitor-1 as a marker of cardiovascular response in professional mountain ultra-marathon runners
  37. Pitfalls of LSD screening assays: comparison of KIMS and CEDIA immunoassays with LC-MS
  38. A two site comparison of two point-of-care activated clotting time systems
  39. Detection of the common South-East Asian β0-thalassemia mutations in samples with borderline HbA2 levels
  40. Biclonal plasma cell myeloma with the simultaneous appearance of both secretory lambda and nonsecretory kappa monoclonal light chains
Sign up now to receive a 20% welcome discount
Institutional Access
How does access work?
Have an idea on how to improve our website?
Please write us.
© 2025 De Gruyter Brill
Downloaded on 27.9.2025 from https://www.degruyterbrill.com/document/doi/10.1515/cclm-2016-0271/html
Scroll to top button