Skip to main content

Advertisement

SpringerLink
Log in

Correlation of MSH2 exonic deletions and protein downregulation with breast cancer biomarkers and outcome in Pakistani women/patients

  • Research Article
  • Published:
Environmental Science and Pollution Research Aims and scope Submit manuscript

Abstract

Mismatch repair (MMR) pathway is one of the underlying mechanisms of predisposition to breast cancer (BC). The present study explored the association of MSH2 exonic deletions, respective survival analysis, protein structure prediction, transcription profiling, and expression analysis with BC risk. Genotyping analysis of 493 BC cases and 387 controls confirmed the association of two MSH2 exonic deletions, i.e., exon 3 (OR:6.4, CI = 3.4–12.1) and 9 (OR:7.8, CI = 4.1–14.8) with BC risk. In order to confirm the phenotypic–genotypic relationship, we have performed MSH2 transcriptomic (p < 0.05) and protein expression analysis (OR:30, CI = 4–230) which further confirmed its downregulation/loss in BC biopsy samples highlighting potential role in the onset of breast carcinogenesis. Additionally, we have presented that MSH2 mutations can alter the expression profile of other BC associated biomarkers like ER, PR, CK–7, GATA–3, and E–cadherin. Subsequently, the effect of exonic deletions on secondary structure of protein has shown missing of beta and alpha helices in their protein products via in-silico analysis. However, loss of exon 3 results in the altered core protein structure leading to dysfunction protein, possible cause of BC development. No association of MSH2 exonic deletions with survival statistics was observed conceivably due to the shorter follow-up time. Thus, our results at genetic, transcriptomic, and proteomic levels confirmed the downregulated MSH2, emphasizing its potential contribution in MMR mechanisms for breast tumorigenesis. In conclusion, MSH2 deficiency may cause breast cancer development and progression.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Availability of data

Data is confidential and stored by removing patient’s identity in excel and SPSS sheets and can be provided upon request.

References

  • Ahmad A (2019) Breast cancer statistics: recent trends. Breast cancer metastasis and drug resistance. Springer, In, pp 1–7

    Google Scholar 

  • Baudhuin LM, Ferber MJ, Winters JL, Steenblock KJ, Swanson RL, French AJ, Butz ML, Thibodeau SN (2005a) Characterization of hMLH1 and hMSH2 gene dosage alterations in Lynch syndrome patients. Gastroenterology 129:846–854

    Article  CAS  Google Scholar 

  • Baudhuin LM, Mai M, French AJ, Kruckeberg KE, Swanson RL, Winters JL, Courteau LK, Thibodeau SN (2005b) Analysis of hMLH1 and hMSH2 gene dosage alterations in hereditary nonpolyposis colorectal cancer patients by novel methods. J Mol Diagn 7:226–235

    Article  CAS  Google Scholar 

  • Britt K (2012) Menarche, menopause, and breast cancer risk. Lancet Oncol 11:1071–1072

    Article  Google Scholar 

  • Chen V, Arendall W, Headd J, Keedy D, Immormino R, Kapral G, Murray L, Richardson J, Richardson D (2010) MolProbity: all-atom contacts and structure validation for proteins and nucleic acids. Acta Cryst D 66:12–21

    Article  CAS  Google Scholar 

  • Cheng AS, Leung SC, Gao D, Burugu S, Anurag M, Ellis MJ, Nielsen TO (2020) Mismatch repair protein loss in breast cancer: clinicopathological associations in a large British Columbia cohort. Breast Cancer Res Treat 179:3–10

    Article  CAS  Google Scholar 

  • Costa RL, Costa-Filho RB, Rosa M, Czerniecki BJ (2017) Occult breast carcinoma presenting as scalp metastasis. Case Rep Oncol 10:992–997

    Article  Google Scholar 

  • DeSantis CE, Bray F, Ferlay J, Lortet-Tieulent J, Anderson BO, Jemal A (2015) International variation in female breast cancer incidence and mortality rates. Cancer Epidem Prev Biomar 24:1495–1506

    Article  Google Scholar 

  • Engel C, Ahadova A, Seppälä T, Aretz S, Bigirwamungu-Bargeman M, Bläker H, Bucksch K, Büttner R, Endris V, Holinski-Feder E (2020) Associations of pathogenic variants in MLH1, MSH2, and MSH6 with risk of colorectal adenomas and tumors and with somatic mutations in patients with Lynch syndrome. Gastroenterology 158:1326–1333

    Article  CAS  Google Scholar 

  • Expert Consultation W (2004) Expert Consultation WHO; WHO Expert Consultation. Appropriate body-mass index for Asian populations and its implications for policy and intervention strategies. Lancet 363:157-163

  • Groothuizen FS, Sixma TK (2016) The conserved molecular machinery in DNA mismatch repair enzyme structures. DNA repair 38:14–23

    Article  CAS  Google Scholar 

  • Guex N, Peitsch M, Schwede T (2009) Automated comparative protein structure modeling with SWISS-MODEL and Swiss-PdbViewer: a historical perspective Electrophoresis. S162–S173, doi 10

  • Hall MJ, Bodor JN, Xiu J, Feldman R, Grothey A, Goldberg RM, Worrilow WM, Hwang JJ, Kim ES, Lenz H-J (2019) Gene-specific features (MLH1, MSH2, MSH6, PMS2) of mismatch repair (MMR) protein expression and somatic mutations (muts), microsatellite instability (MSI) and tumor mutational burden (TMB) in MSI-H and MMR-mutated tumor genomic profiles (TGPs). In. American Society of Clinical Oncology

  • Hobeika E, Armouti M, Kala HS, Stocco C (2020) Ovarian hormones. Hormonal signaling in Biology and Medicine. Elsevier, In, pp 565–583

    Google Scholar 

  • Hsieh Y-C, Cho E-C, Tu S-H, Wu C-H, Hung C-S, Hsieh M-C, Su C-T, Liu Y-R, Lee C-H, Ho Y-S (2017) Msh2 rs2303425 polymorphism is associated with early-onset breast cancer in Taiwan. Ann Surg Oncol 24:603–610

    Article  Google Scholar 

  • Jaballah-Gabteni A, Tounsi H, Kabbage M, Hamdi Y, Elouej S, Ayed IB, Medhioub M, Mahmoudi M, Dallali H, Yaiche H (2019) Identification of novel pathogenic MSH2 mutation and new DNA repair genes variants: investigation of a Tunisian Lynch syndrome family with discordant twins. J Transl Med 17:212

    Article  CAS  Google Scholar 

  • Kadyrov FA, Dzantiev L, Constantin N, Modrich P (2006) Endonucleolytic function of MutLα in human mismatch repair. Cell 126:297–308

    Article  CAS  Google Scholar 

  • Kappil M, Terry MB, Delgado-Cruzata L, Liao Y, Santella RM (2016) Mismatch repair polymorphisms as markers of breast cancer prevalence in the breast cancer family registry. Anticancer Res 36:4437–4441

    Article  CAS  Google Scholar 

  • Kelley L, Mezulis S, Yates C, Wass M, Sternberg M (2015) The Phyre2 web portal for protein modeling, prediction and analysis. Nat Protoc 10:845–858

    Article  CAS  Google Scholar 

  • Letunic I, Doerks T, Bork P (2015) SMART: recent updates, new developments and status in 2015. Nucleic Acids Res 43:D257–D260

    Article  CAS  Google Scholar 

  • Lipkin SM, Wang V, Jacoby R, Banerjee-Basu S, Baxevanis AD, Lynch HT, Elliott RM, Collins FS (2000) MLH3: a DNA mismatch repair gene associated with mammalian microsatellite instability. Nat Genet 24:27–35

    Article  CAS  Google Scholar 

  • Lu F-I, Gilks CB, Mulligan A-M, Ryan P, Allo G, Sy K, Shaw PA, Pollett A, Clarke BA (2012) Prevalence of loss of expression of DNA mismatch repair proteins in primary epithelial ovarian tumors. Int J Gynecol Pathol 31:524–531

    Article  CAS  Google Scholar 

  • Malik SS, Mubarik S, Masood N, Khadim MT (2018) An insight into clinical outcome of XPG polymorphisms in breast cancer. Mol Biol Rep 45:2369–2375

    Article  CAS  Google Scholar 

  • Malik SS, Baig M, Khan MB, Masood N (2019) Survival analysis of breast cancer patients with different treatments: a multi-centric clinicopathological study. JPMA

  • Malik SS, Zia A, Mubarik S, Masood N, Rashid S, Sherrard A, Khan MB, Khadim MT (2020) Correlation of MLH1 polymorphisms, survival statistics, in silico assessment and gene downregulation with clinical outcomes among breast cancer cases. Mol Biol Rep 47:683–692

    Article  CAS  Google Scholar 

  • Mavaddat N, Pharoah PD, Michailidou K, Tyrer J, Brook MN, Bolla MK, Wang Q, Dennis J, Dunning AM, Shah M (2015) Prediction of breast cancer risk based on profiling with common genetic variants. JNCI: Journal of the National Cancer Institute 107

  • Meng X, Song S, Z-f J, Sun B, Wang T, Zhang S, Wu S (2016) Receptor conversion in metastatic breast cancer: a prognosticator of survival. Oncotarget 7:71887–71903

    Article  Google Scholar 

  • Miettinen M, Cue PAM, Sarlomo-Rikala M, Rys J, Czapiewski P, Wazny K, Langfort R, Waloszczyk P, Biernat W, Lasota J (2014) GATA 3–a multispecific but potentially useful marker in surgical pathology–a systematic analysis of 2500 epithelial and non-epithelial tumors. Am J Surg Pathol 38:13–22

    Article  Google Scholar 

  • Miki Y, Swensen J, Shattuck-Eidens D, Futreal PA, Harshman K, Tavtigian S, Liu Q, Cochran C, Bennett LM, Ding W (1994) A strong candidate for the breast and ovarian cancer susceptibility gene BRCA1. Science 266:66–71

    Article  CAS  Google Scholar 

  • Molaei M, Mansoori BK, Ghiasi S, Khatami F, Attarian H, Zali M (2010) Colorectal cancer in Iran: immunohistochemical profiles of four mismatch repair proteins. Int J Color Dis 25:63–69

    Article  Google Scholar 

  • Mubarik S, Malik SS, Wang Z, Li C, Fawad M, Yu C (2019) Recent insights into breast cancer incidence trends among four Asian countries using age-period-cohort model. Cancer Manag Res 11:8145–8155

    Article  Google Scholar 

  • Naidu MD, Mason JM, Pica RV, Fung H, Peña LA (2010) Radiation resistance in glioma cells determined by DNA damage repair activity of Ape1/Ref-1. J Radiat Res 51:393–404

    Article  CAS  Google Scholar 

  • Owczarzy R, Tataurov AV, Wu Y, Manthey JA, McQuisten KA, Almabrazi HG, Pedersen KF, Lin Y, Garretson J, McEntaggart NO (2008) IDT SciTools: a suite for analysis and design of nucleic acid oligomers. Nucleic Acids Res 36:W163–W169

    Article  CAS  Google Scholar 

  • Park YJ, Shin K-H, Park J-G (2000) Risk of gastric cancer in hereditary nonpolyposis colorectal cancer in Korea. Clin Cancer Res 6:2994–2998

    CAS  Google Scholar 

  • Poulogiannis G, Frayling IM, Arends MJ (2010) DNA mismatch repair deficiency in sporadic colorectal cancer and Lynch syndrome. Histopathology 56:167–179

    Article  Google Scholar 

  • Pritchard CC, Morrissey C, Kumar A, Zhang X, Smith C, Coleman I, Salipante SJ, Milbank J, Yu M, Grady WM, Tait JF, Corey E, Vessella RL, Walsh T, Shendure J, Nelson PS (2014) Complex MSH2 and MSH6 mutations in hypermutated microsatellite unstable advanced prostate cancer. Nat Commun 5:4988

    Article  CAS  Google Scholar 

  • Rashid MU, Naeemi H, Muhammad N, Loya A, Lubiński J, Jakubowska A, Yusuf MA (2019) Prevalence and spectrum of MLH1, MSH2, and MSH6 pathogenic germline variants in Pakistani colorectal cancer patients. Hered Cancer Clin Pract 17:29

    Article  CAS  Google Scholar 

  • Rhees J, Arnold M, Boland CR (2014) Inversion of exons 1–7 of the MSH2 gene is a frequent cause of unexplained Lynch syndrome in one local population. Familial Cancer 13:219–225

    Article  CAS  Google Scholar 

  • da Silva Calixto P, Lopes OS, dos Santos Maia M, Herrero SST, Longui CA, Melo CGF, de Carvalho Filho IR, Soares LF, de Medeiros AC, Delatorre P, khayat AS, Burbano RR, Lima EM (2017) Single-nucleotide polymorphisms of the MSH2 and MLH1 genes, potential molecular markers for susceptibility to the development of basal cell carcinoma in the Brazilian population. Pathol Oncol Res

  • Singhai R, Patil VW, Jaiswal SR, Patil SD, Tayade MB, Patil AV (2011) E-Cadherin as a diagnostic biomarker in breast cancer. N Am J Med Sci 3:227

    Article  Google Scholar 

  • Smith MJ, Urquhart JE, Harkness EF, Miles EK, Bowers NL, Byers HJ, Bulman M, Gokhale C, Wallace AJ, Newman WG (2016) The contribution of whole gene deletions and large rearrangements to the mutation spectrum in inherited tumor predisposing syndromes. Hum Mutat 37:250–256

    Article  CAS  Google Scholar 

  • Stella A, Surdo N, Lastella P, Barana D, Oliani C, Tibiletti M, Viel A, Natale C, Piepoli A, Marra G (2007) Germline novel MSH2 deletions and a founder MSH2 deletion associated with anticipation effects in HNPCC. Clin Genet 71:130–139

    Article  CAS  Google Scholar 

  • Thiffault I, Hamel N, Pal T, McVety S, Marcus V, Farber D, Cowie S, Deschenes J, Meschino W, Odefrey F (2004) Germline truncating mutations in both MSH2 and BRCA2 in a single kindred. Br J Cancer 90:483–491

    Article  CAS  Google Scholar 

  • Tung N, Lin NU, Kidd J, Allen BA, Singh N, Wenstrup RJ, Hartman A-R, Winer EP, Garber JE (2016) Frequency of germline mutations in 25 cancer susceptibility genes in a sequential series of patients with breast cancer. J Clin Oncol 34:1460–1468

    Article  CAS  Google Scholar 

  • Verma R, Agarwal AK, Sakhuja P, Sharma PC (2019) Microsatellite instability in mismatch repair and tumor suppressor genes and their expression profiling provide important targets for the development of biomarkers in gastric cancer. Gene 710:48–58

    Article  CAS  Google Scholar 

  • Watson P, Vasen HF, Mecklin JP, Bernstein I, Aarnio M, Järvinen HJ, Myrhøj T, Sunde L, Wijnen JT, Lynch HT (2008) The risk of extra-colonic, extra-endometrial cancer in the Lynch syndrome. Int J Cancer 123:444–449

    Article  CAS  Google Scholar 

  • Westenend PJ, Schütte R, Hoogmans MM, Wagner A, Dinjens WN (2005) Breast cancer in an MSH2 gene mutation carrier. Hum Pathol 36:1322–1326

    Article  CAS  Google Scholar 

  • Wooster R, Neuhausen SL, Mangion J, Quirk Y, Ford D, Collins N, Nguyen K, Seal S, Tran T, Averill D (1994) Localization of a breast cancer susceptibility gene, BRCA2, to chromosome 13q12-13. Science 265:2088–2090

    Article  CAS  Google Scholar 

  • Wu B, Ji W, Liang S, Ling C, You Y, Xu L, Zhong M-E, Xiao Y, Qiu H-Z, Lu J-Y, Banerjee S (2017) A novel heterozygous germline deletion in MSH2 gene in a five generation Chinese family with Lynch syndrome. Oncotarget 8:55194–55203

    Article  Google Scholar 

  • Xiang T-X, Yuan Y, Li L-L, Wang Z-H, Dan L-Y, Chen Y, Ren G-S, Tao Q (2013) Aberrant promoter CpG methylation and its translational applications in breast cancer. Chin J Cancer 32:12–20

    Article  CAS  Google Scholar 

  • Zhu M, Chen HM, Wang YP (2013) Missense mutations of MLH1 and MSH2 genes detected in patients with gastrointestinal cancer are associated with exonic splicing enhancers and silencers. Oncol Lett 5:1710–1718

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We would like to thank all the patients and their attendees for their support. We also like to thank Armed Forces Institute of Pathology (AFIP), for providing lab facility.

Author information

Author notes

  1. Ayesha Aftab and Ranjha Khan contributed equally to this work.

Authors and Affiliations

  1. Department of Zoology, University of Gujrat, Gujrat, Pakistan

    Saima Shakil Malik

  2. Microbiology & Biotechnology Research Lab., Fatima Jinnah Women University, The Mall Rawalpindi, Rawalpindi, Punjab, Pakistan

    Saima Shakil Malik & Nosheen Masood

  3. Department of Epidemiology and Biostatistics, School of Health Sciences, Wuhan University, Wuhan, 430071, Hubei, China

    Sumaira Mubarik

  4. Department of Biological Sciences, International Islamic University, Sector H10, Islamabad, Pakistan

    Ayesha Aftab

  5. The CAS Key Laboratory of Innate Immunity and Chronic Diseases, School of Life Sciences, University of Science and Technology of China, Huangshan Road, Hefei, 230027, Anhui, China

    Ranjha Khan

  6. Department of Histopathology, Armed Forces Institute of Pathology, Rawalpindi, Pakistan

    Muhammad Asif

  7. Breast Clinic, Combined Military Hospital, Rawalpindi, Pakistan

    Razia Bano

Authors
  1. Saima Shakil Malik
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sumaira Mubarik
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ayesha Aftab
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ranjha Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Nosheen Masood
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Muhammad Asif
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Razia Bano
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

SSM—Conceptualization, sample collection, lab experimentation, write up; SM—Statistical analysis and interpretation, AA—In-silico analysis, interpretation and write up; RK—Experiment design, interpretation, and write up; NM—Conceptualization, supervision, experiment design, proof reading; MA—is a histopathologist, supervises and executes expression studies; RB—is a surgeon—tissue sampling, handling, and preservation

Corresponding author

Correspondence to Saima Shakil Malik.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethics approval and consent to participate

Ethical approval was obtained from all the related hospital and sampling was done after taking written informed consent from the patients as the study involves human participants. Ethical approval letters and sample questionnaire can be provided if needed.

Additional information

Responsible editor: Lotfi Aleya

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

ESM 1

(DOCX 933 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Malik, S.S., Mubarik, S., Aftab, A. et al. Correlation of MSH2 exonic deletions and protein downregulation with breast cancer biomarkers and outcome in Pakistani women/patients. Environ Sci Pollut Res 28, 3066–3077 (2021). https://doi.org/10.1007/s11356-020-10717-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11356-020-10717-z

Keywords

Search

Navigation