FHIT promoter methylation status, low protein and high mRNA levels in patients with non-small cell lung cancer

Czarnecka,Karolina  H.; Migdalska-Sęk,Monika; Domańska,Daria; Pastuszak-Lewandoska,Dorota; Dutkowska,Agata; Kordiak,Jacek; Nawrot,Ewa; Kiszałkiewicz,Justyna; Antczak,Adam; Brzeziańska-Lasota,Ewa

doi:10.3892/ijo.2016.3610

FHIT promoter methylation status, low protein and high mRNA levels in patients with non-small cell lung cancer

Authors:
- Karolina H. Czarnecka
- Monika Migdalska-Sęk
- Daria Domańska
- Dorota Pastuszak-Lewandoska
- Agata Dutkowska
- Jacek Kordiak
- Ewa Nawrot
- Justyna Kiszałkiewicz
- Adam Antczak
- Ewa Brzeziańska-Lasota
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Affiliations: Department of Molecular Bases of Medicine, Medical University of Lodz, Lodz, Poland, Department of General and Oncological Pulmonology, Medical University of Lodz, Lodz, Poland, Department of Chest Surgery, General and Oncological Surgery University Hospital No. 2, Medical University of Lodz, Lodz, Poland
Published online on: July 6, 2016 https://doi.org/10.3892/ijo.2016.3610
Pages: 1175-1184

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Abstract

FHIT is a tumor suppressor gene that is frequently silenced in non-small cell lung cancer (NSCLC) and also in preneoplastic lesions. Promoter hypermethylation was previously observed in NSCLC, and its epigenetic silencing, observed on mRNA or protein level, was proposed to predict NSCLC outcome. In the present study we evaluated the relationship between FHIT expression on mRNA level and promoter methylation, or immunoexpression level. The aim of this study was to analyze the usefulness of FHIT as early differentiating biomarker in NSCLC patients. Lung tissue specimens were obtained from 59 patients with diagnosed NSCLC (SCC=34, AC=20, LCC=5). FHIT promoter methylation was assessed in methylation-specific PCR. Relative expression analysis of FHIT was performed in real-time PCR (qPCR) and protein immunoexpression by ELISA assay. Significant differences in FHIT expression between NSCLC histopathological groups (SCC, AC, LCC) were observed (p=0.000009), with the lowest level in SCC. FHIT expression was significantly higher (p=0.034) in men vs. women. Methylated FHIT alleles were present both in NSCLC and control specimens. Mean MI value was higher in control tissue vs. neoplasm, and in men vs. women and it increased with patient age. Significant increase in MI level was observed in N0 group vs. N1 and N2, according to the TNM staging (p=0.0073). Differences in FHIT expression levels between AC, LCC and SCC indicated the usefulness of this gene as a diagnostic marker for NSCLC subtype differentiation. FHIT promoter hypermethylation both in cancer and control tissue indicated the presence of epigenetic alterations in early stage of NSCLC development. Differences in gene promoter methylation between cancer patients with and without node infiltration might be considered as a prognostic marker. Significantly lower FHIT protein immunoexpression was revealed in the group with long and intense history of smoking assessed as PYs (PY<40 vs. PY≥40, p=0.01). These results suggest the need of further study on FHIT as a potential biomarker.

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1	Wojciechowska U, Didkowska J and Zatoński W: Malignant neoplasms. Cancer in Poland in 2012. Oncology Centre - Marie Curie Institute, . Warszawa: pp. 11–25. 2012
2	Ferlay J, Shin HR, Bray F, Forman D, Mathers C and Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 127:2893–2917. 2010. View Article : Google Scholar
3	Travis WD, Travis LB and Devesa SS: Lung cancer. Cancer. 75(Suppl): 191–202. 1995. View Article : Google Scholar : PubMed/NCBI
4	Travis WD, Brambilla E, Müller-Hermelink HK and Harris CC: Tumours of the lung. WHO Classification Pathology and Genetics of Tumours of the Lung, Pleura, Thymus and Heart. IARC Press; pp. 9–122. 2004
5	Kathuria H, Gesthalter Y, Spira A, Brody JS and Steiling K: Updates and controversies in the rapidly evolving field of lung cancer screening, early detection, and chemoprevention. Cancers (Basel). 6:1157–1179. 2014. View Article : Google Scholar
6	Zabarovsky ER, Lerman MI and Minna JD: Tumor suppressor genes on chromosome 3p involved in the pathogenesis of lung and other cancers. Oncogene. 21:6915–6935. 2002. View Article : Google Scholar : PubMed/NCBI
7	Ohta M, Inoue H, Cotticelli MG, Kastury K, Baffa R, Palazzo J, Siprashvili Z, Mori M, McCue P, Druck T, et al: The FHIT gene, spanning the chromosome 3p14.2 fragile site and renal carcinoma-associated t(3;8) breakpoint, is abnormal in digestive tract cancers. Cell. 84:587–597. 1996. View Article : Google Scholar : PubMed/NCBI
8	Semba S, Trapasso F, Fabbri M, McCorkell KA, Volinia S, Druck T, Iliopoulos D, Pekarsky Y, Ishii H, Garrison PN, et al: Fhit modulation of the Akt-survivin pathway in lung cancer cells: Fhit-tyrosine 114 (Y114) is essential. Oncogene. 25:2860–2872. 2006. View Article : Google Scholar : PubMed/NCBI
9	Weiske J, Albring KF and Huber O: The tumor suppressor Fhit acts as a repressor of beta-catenin transcriptional activity. Proc Natl Acad Sci USA. 104:20344–20349. 2007. View Article : Google Scholar : PubMed/NCBI
10	Siprashvili Z, Sozzi G, Barnes LD, McCue P, Robinson AK, Eryomin V, Sard L, Tagliabue E, Greco A, Fusetti L, et al: Replacement of Fhit in cancer cells suppresses tumorigenicity. Proc Natl Acad Sci USA. 94:13771–13776. 1997. View Article : Google Scholar
11	Roz L, Gramegna M, Ishii H, Croce CM and Sozzi G: Restoration of fragile histidine triad (FHIT) expression induces apoptosis and suppresses tumorigenicity in lung and cervical cancer cell lines. Proc Natl Acad Sci USA. 99:3615–3620. 2002. View Article : Google Scholar : PubMed/NCBI
12	Fong KM, Biesterveld EJ, Virmani A, Wistuba I, Sekido Y, Bader SA, Ahmadian M, Ong ST, Rassool FV, Zimmerman PV, et al: FHIT and FRA3B 3p14.2 allele loss are common in lung cancer and preneoplastic bronchial lesions and are associated with cancer-related FHIT cDNA splicing aberrations. Cancer Res. 57:2256–2267. 1997.PubMed/NCBI
13	Pichiorri F, Okumura H, Nakamura T, Garrison PN, Gasparini P, Suh SS, Druck T, McCorkell KA, Barnes LD, Croce CM, et al: Correlation of fragile histidine triad (Fhit) protein structural features with effector interactions and biological functions. J Biol Chem. 284:1040–1049. 2009. View Article : Google Scholar :
14	Saldivar JC, Bene J, Hosseini SA, Miuma S, Horton S, Heerem NA and Huebner K: Characterization of the role of Fhit in suppression of DNA damage. Adv Biol Regul. 53:77–85. 2013. View Article : Google Scholar :
15	Wang HL, Zhou PY, Liu P and Zhang Y: Abnormal FHIT protein expression may be correlated with poor prognosis in gastric cancer: A meta-analysis. Tumour Biol. 35:6815–6821. 2014. View Article : Google Scholar : PubMed/NCBI
16	Joannes A, Bonnomet A, Bindels S, Polette M, Gilles C, Burlet H, Cutrona J, Zahm JM, Birembaut P and Nawrocki-Raby B: Fhit regulates invasion of lung tumor cells. Oncogene. 29:1203–1213. 2010. View Article : Google Scholar
17	Toledo G, Sola JJ, Lozano MD, Soria E and Pardo J: Loss of FHIT protein expression is related to high proliferation, low apoptosis and worse prognosis in non-small-cell lung cancer. Mod Pathol. 17:440–448. 2004. View Article : Google Scholar : PubMed/NCBI
18	Kim JS, Kim JW, Han J, Shim YM, Park J and Kim DH: Cohypermethylation of p16 and FHIT promoters as a prognostic factor of recurrence in surgically resected stage I non-small cell lung cancer. Cancer Res. 66:4049–4054. 2006. View Article : Google Scholar : PubMed/NCBI
19	American Joint Committee on Cancer Staging according to the IASLC Staging Project. 7th edition. Cancer. 2010, http://cancerstaging.org/references-tools/quickreferences/documents/lungmedium.pdf.
20	Herman JG, Graff JR, Myöhänen S, Nelkin BD and Baylin SB: Methylation-specific PCR: A novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA. 93:9821–9826. 1996. View Article : Google Scholar : PubMed/NCBI
21	Feltus FA, Lee EK, Costello JF, Plass C and Vertino PM: Predicting aberrant CpG island methylation. Proc Natl Acad Sci USA. 100:12253–12258. 2003. View Article : Google Scholar : PubMed/NCBI
22	Li LC and Dahiya R: MethPrimer: Designing primers for methylation PCRs. Bioinformatics. 18:1427–1431. 2002. View Article : Google Scholar : PubMed/NCBI
23	Sozzi G, Tornielli S, Tagliabue E, Sard L, Pezzella F, Pastorino U, Minoletti F, Pilotti S, Ratcliffe C, Veronese ML, et al: Absence of Fhit protein in primary lung tumors and cell lines with FHIT gene abnormalities. Cancer Res. 57:5207–5212. 1997.PubMed/NCBI
24	Sozzi G, Sard L, De Gregorio L, Marchetti A, Musso K, Buttitta F, Tornielli S, Pellegrini S, Veronese ML, Manenti G, et al: Association between cigarette smoking and FHIT gene alterations in lung cancer. Cancer Res. 57:2121–2123. 1997.PubMed/NCBI
25	Sozzi G, Pastorino U, Moiraghi L, Tagliabue E, Pezzella F, Ghirelli C, Tornielli S, Sard L, Huebner K, Pierotti MA, et al: Loss of FHIT function in lung cancer and preinvasive bronchial lesions. Cancer Res. 58:5032–5037. 1998.PubMed/NCBI
26	Zhao P, Li XY and Chen LZ: Loss of fragile histidine triad expression and metastasis in breast cancer. Ai Zheng. 21:668–670. 2002.(In Chinese). PubMed/NCBI
27	Bekar A, Ceçener G, Tunca B, Guler G, Egeli U and Tolunay S: Investigation of mutations and expression of the FHIT gene in Turkish patients with brain metastases derived from non-small cell lung cancer. Tumori. 93:604–607. 2007.
28	Suh SS, Yoo JY, Cui R, Kaur B, Huebner K, Lee TK, Aqeilan RI and Croce CM: FHIT suppresses epithelial-mesenchymal transition (EMT) and metastasis in lung cancer through modulation of microRNAs. PLoS Genet. 10:e10046522014. View Article : Google Scholar : PubMed/NCBI
29	Song X, Tian Z, Wang S, Peng Z and Feng J: Restoration of fragile histidine triad (FHIT) expression inhibits cell growth and induces apoptosis in cutaneous T-cell lymphoma cell line. Cancer Invest. 28:1019–1023. 2010. View Article : Google Scholar : PubMed/NCBI
30	Zanesi N, Fidanza V, Fong LY, Mancini R, Druck T, Valtieri M, Rüdiger T, McCue PA, Croce CM and Huebner K: The tumor spectrum in FHIT-deficient mice. Proc Natl Acad Sci USA. 98:10250–10255. 2001. View Article : Google Scholar : PubMed/NCBI
31	Geradts J, Fong KM, Zimmerman PV and Minna JD: Loss of Fhit expression in non-small-cell lung cancer: Correlation with molecular genetic abnormalities and clinicopathological features. Br J Cancer. 82:1191–1197. 2000. View Article : Google Scholar : PubMed/NCBI
32	Pavelić K, Krizanac S, Cacev T, Hadzija MP, Radosević S, Crnić I, Levanat S and Kapitanović S: Aberration of FHIT gene is associated with increased tumor proliferation and decreased apoptosis-clinical evidence in lung and head and neck carcinomas. Mol Med. 7:442–453. 2001.
33	Feng Q, Hawes SE, Stern JE, Wiens L, Lu H, Dong ZM, Jordan CD, Kiviat NB and Vesselle H: DNA methylation in tumor and matched normal tissues from non-small cell lung cancer patients. Cancer Epidemiol Biomarkers Prev. 17:645–654. 2008. View Article : Google Scholar : PubMed/NCBI
34	Tomizawa Y, Nakajima T, Kohno T, Saito R, Yamaguchi N and Yokota J: Clinicopathological significance of Fhit protein expression in stage I non-small cell lung carcinoma. Cancer Res. 58:5478–5483. 1998.PubMed/NCBI
35	Tseng JE, Kemp BL, Khuri FR, Kurie JM, Lee JS, Zhou X, Liu D, Hong WK and Mao L: Loss of Fhit is frequent in stage I non-small cell lung cancer and in the lungs of chronic smokers. Cancer Res. 59:4798–4803. 1999.PubMed/NCBI
36	Lindskog C, Edlund K, Mattsson JS and Micke P: Immunohistochemistry-based prognostic biomarkers in NSCLC: Novel findings on the road to clinical use? Expert Rev Mol Diagn. 15:471–490. 2015. View Article : Google Scholar : PubMed/NCBI
37	Marchetti A, Pellegrini S, Bertacca G, Buttitta F, Gaeta P, Carnicelli V, Nardini V, Griseri P, Chella A, Angeletti CA, et al: FHIT and p53 gene abnormalities in bronchioloalveolar carcinomas. Correlations with clinicopathological data and K-ras mutations. J Pathol. 184:240–246. 1998. View Article : Google Scholar : PubMed/NCBI