Background Although early-stage non-small-cell lung cancer (NSCLC) is considered a potentially curable disease following complete resection, patients have a wide spectrum of survival according to stage (IB, II, IIIA). Within each stage, gene expression profiles can identify patients with a higher risk of recurrence. We hypothesized that altered mRNA expression in nine genes could help to predict disease outcome: excision repair cross-complementing 1 (ERCC1), myeloid zinc finger 1 (MZF1) and Twist1 (which regulate N-cadherin expression), ribonucleotide reductase subunit M1 (RRM1), thioredoxin-1 (TRX1), tyrosyl-DNA phosphodiesterase (Tdp1), nuclear factor of activated T cells (NFAT), BRCA1, and the human homolog of yeast budding uninhibited by benzimidazole (BubR1). Methodology and Principal Findings We performed real-time quantitative polymerase chain reaction (RT-QPCR) in frozen lung cancer tissue specimens from 126 chemonaive NSCLC patients who had undergone surgical resection and evaluated the association between gene expression levels and survival. For validation, we used paraffin-embedded specimens from 58 other NSCLC patients. A strong inter-gene correlation was observed between expression levels of all genes except NFAT. A Cox proportional hazards model indicated that along with disease stage, BRCA1 mRNA expression significantly correlated with overall survival (hazard ratio [HR], 1.98 [95% confidence interval (CI), 1.11-6]; P = 0.02). In the independent cohort of 58 patients, BRCA1 mRNA expression also significantly correlated with survival (HR, 2.4 [95%CI, 1.01-5.92]; P = 0.04). Conclusions Overexpression of BRCA1 mRNA was strongly associated with poor survival in NSCLC patients, and the validation of this finding in an independent data set further strengthened this association. Since BRCA1 mRNA expression has previously been linked to differential sensitivity to cisplatin and antimicrotubule drugs, BRCA1 mRNA expression may provide additional information for customizing adjuvant antimicrotubule-based chemotherapy, especially in stage IB, where the role of adjuvant chemotherapy has not been clearly demonstrated.
References
[1]
Ferlay J, Autier P, Boniol M, Heanue M, Colombet M, et al. (2007) Estimates of the cancer incidence and mortality in Europe in 2006. Ann Oncol 18: 581–592.
[2]
Strauss GM (2005) Adjuvant chemotherapy of lung cancer: methodologic issues and therapeutic advances. Hematol Oncol Clin North Am 19: 263–281, vi.
[3]
Douillard JY, Rosell R, De Lena M, Carpagnano F, Ramlau R, et al. (2006) Adjuvant vinorelbine plus cisplatin versus observation in patients with completely resected stage IB-IIIA non-small-cell lung cancer (Adjuvant Navelbine International Trialist Association [ANITA]): a randomised controlled trial. Lancet Oncol 7: 719–727.
[4]
Parmigiani G, Garrett-Mayer ES, Anbazhagan R, Gabrielson E (2004) A cross-study comparison of gene expression studies for the molecular classification of lung cancer. Clin Cancer Res 10: 2922–2927.
[5]
Potti A, Mukherjee S, Petersen R, Dressman HK, Bild A, et al. (2006) A genomic strategy to refine prognosis in early-stage non-small-cell lung cancer. N Engl J Med 355: 570–580.
[6]
Lu Y, Lemon W, Liu PY, Yi Y, Morrison C, et al. (2006) A gene expression signature predicts survival of patients with stage I non-small cell lung cancer. PLoS Med 3: e467.
[7]
Chen HY, Yu SL, Chen CH, Chang GC, Chen CY, et al. (2007) A five-gene signature and clinical outcome in non-small-cell lung cancer. N Engl J Med 356: 11–20.
[8]
Nevins JR, Potti A (2007) Mining gene expression profiles: expression signatures as cancer phenotypes. Nat Rev Genet.
[9]
Furuta T, Ueda T, Aune G, Sarasin A, Kraemer KH, et al. (2002) Transcription-coupled nucleotide excision repair as a determinant of cisplatin sensitivity of human cells. Cancer Res 62: 4899–4902.
[10]
Lord RV, Brabender J, Gandara D, Alberola V, Camps C, et al. (2002) Low ERCC1 expression correlates with prolonged survival after cisplatin plus gemcitabine chemotherapy in non-small cell lung cancer. Clin Cancer Res 8: 2286–2291.
[11]
Taron M, Rosell R, Felip E, Mendez P, Souglakos J, et al. (2004) BRCA1 mRNA expression levels as an indicator of chemoresistance in lung cancer. Hum Mol Genet 13: 2443–2449.
[12]
Bae I, Rih JK, Kim HJ, Kang HJ, Haddad B, et al. (2005) BRCA1 regulates gene expression for orderly mitotic progression. Cell Cycle 4: 1641–1666.
[13]
Chabalier C, Lamare C, Racca C, Privat M, Valette A, et al. (2006) BRCA1 downregulation leads to premature inactivation of spindle checkpoint and confers paclitaxel resistance. Cell Cycle 5: 1001–1007.
[14]
Shichiri M, Yoshinaga K, Hisatomi H, Sugihara K, Hirata Y (2002) Genetic and epigenetic inactivation of mitotic checkpoint genes hBUB1 and hBUBR1 and their relationship to survival. Cancer Res 62: 13–17.
[15]
Hromas R, Collins SJ, Hickstein D, Raskind W, Deaven LL, et al. (1991) A retinoic acid-responsive human zinc finger gene, MZF-1, preferentially expressed in myeloid cells. J Biol Chem 266: 14183–14187.
[16]
Yan QW, Reed E, Zhong XS, Thornton K, Guo Y, et al. (2006) MZF1 possesses a repressively regulatory function in ERCC1 expression. Biochem Pharmacol 71: 761–771.
[17]
Le Mee S, Fromigue O, Marie PJ (2005) Sp1/Sp3 and the myeloid zinc finger gene MZF1 regulate the human N-cadherin promoter in osteoblasts. Exp Cell Res 302: 129–142.
[18]
Rosell R, Scagliotti G, Danenberg KD, Lord RV, Bepler G, et al. (2003) Transcripts in pretreatment biopsies from a three-arm randomized trial in metastatic non-small-cell lung cancer. Oncogene 22: 3548–3553.
[19]
Zheng Z, Chen T, Li X, Haura E, Sharma A, et al. (2007) DNA synthesis and repair genes RRM1 and ERCC1 in lung cancer. N Engl J Med 356: 800–808.
[20]
Rosell R, Felip E, Taron M, Majo J, Mendez P, et al. (2004) Gene expression as a predictive marker of outcome in stage IIB-IIIA-IIIB non-small cell lung cancer after induction gemcitabine-based chemotherapy followed by resectional surgery. Clin Cancer Res 10: 4215s–4219s.
[21]
Kakolyris S, Giatromanolaki A, Koukourakis M, Powis G, Souglakos J, et al. (2001) Thioredoxin expression is associated with lymph node status and prognosis in early operable non-small cell lung cancer. Clin Cancer Res 7: 3087–3091.
[22]
Yoshida T, Nakamura H, Masutani H, Yodoi J (2005) The involvement of thioredoxin and thioredoxin binding protein-2 on cellular proliferation and aging process. Ann N Y Acad Sci 1055: 1–12.
[23]
Interthal H, Chen HJ, Kehl-Fie TE, Zotzmann J, Leppard JB, et al. (2005) SCAN1 mutant Tdp1 accumulates the enzyme–DNA intermediate and causes camptothecin hypersensitivity. Embo J 24: 2224–2233.
[24]
Barthelmes HU, Habermeyer M, Christensen MO, Mielke C, Interthal H, et al. (2004) TDP1 overexpression in human cells counteracts DNA damage mediated by topoisomerases I and II. J Biol Chem 279: 55618–55625.
[25]
Liu C, Zhou S, Begum S, Sidransky D, Westra WH, et al. (2007) Increased expression and activity of repair genes TDP1 and XPF in non-small cell lung cancer. Lung Cancer 55: 303–311.
[26]
Yang J, Mani SA, Donaher JL, Ramaswamy S, Itzykson RA, et al. (2004) Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasis. Cell 117: 927–939.
[27]
Alexander NR, Tran NL, Rekapally H, Summers CE, Glackin C, et al. (2006) N-cadherin gene expression in prostate carcinoma is modulated by integrin-dependent nuclear translocation of Twist1. Cancer Res 66: 3365–3369.
[28]
Yoeli-Lerner M, Yiu GK, Rabinovitz I, Erhardt P, Jauliac S, et al. (2005) Akt blocks breast cancer cell motility and invasion through the transcription factor NFAT. Mol Cell 20: 539–550.
[29]
Jauliac S, Lopez-Rodriguez C, Shaw LM, Brown LF, Rao A, et al. (2002) The role of NFAT transcription factors in integrin-mediated carcinoma invasion. Nat Cell Biol 4: 540–544.
[30]
Mountain CF (1997) Revisions in the International System for Staging Lung Cancer. Chest 111: 1710–1717.
[31]
Lausen B, Schumacher M (1992) Maximally selected rank statistics. Biometrics 48: 73–85.
[32]
Endoh H, Tomida S, Yatabe Y, Konishi H, Osada H, et al. (2004) Prognostic model of pulmonary adenocarcinoma by expression profiling of eight genes as determined by quantitative real-time reverse transcriptase polymerase chain reaction. J Clin Oncol 22: 811–819.
[33]
Quinn JE, Kennedy RD, Mullan PB, Gilmore PM, Carty M, et al. (2003) BRCA1 functions as a differential modulator of chemotherapy-induced apoptosis. Cancer Res 63: 6221–6228.
[34]
Wisnivesky JP, Yankelevitz D, Henschke CI (2004) The effect of tumor size on curability of stage I non-small cell lung cancers. Chest 126: 761–765.
[35]
Potapova O, Haghighi A, Bost F, Liu C, Birrer MJ, et al. (1997) The Jun kinase/stress-activated protein kinase pathway functions to regulate DNA repair and inhibition of the pathway sensitizes tumor cells to cisplatin. J Biol Chem 272: 14041–14044.
[36]
Zhao R, Rabo YB, Egyhazi S, Andersson A, Edgren MR, et al. (1995) Apoptosis and c-jun induction by cisplatin in a human melanoma cell line and a drug-resistant daughter cell line. Anticancer Drugs 6: 657–668.
[37]
Moorehead RA, Singh G (2000) Influence of the proto-oncogene c-fos on cisplatin sensitivity. Biochem Pharmacol 59: 337–345.
[38]
Nakamura H (2004) Thioredoxin as a key molecule in redox signaling. Antioxid Redox Signal 6: 15–17.
[39]
Welsh SJ, Bellamy WT, Briehl MM, Powis G (2002) The redox protein thioredoxin-1 (Trx-1) increases hypoxia-inducible factor 1alpha protein expression: Trx-1 overexpression results in increased vascular endothelial growth factor production and enhanced tumor angiogenesis. Cancer Res 62: 5089–5095.
[40]
Rice JC, Massey-Brown KS, Futscher BW (1998) Aberrant methylation of the BRCA1 CpG island promoter is associated with decreased BRCA1 mRNA in sporadic breast cancer cells. Oncogene 17: 1807–1812.
[41]
Marsit CJ, Liu M, Nelson HH, Posner M, Suzuki M, et al. (2004) Inactivation of the Fanconi anemia/BRCA pathway in lung and oral cancers: implications for treatment and survival. Oncogene 23: 1000–1004.
[42]
Grushko TA, Dignam JJ, Das S, Blackwood AM, Perou CM, et al. (2004) MYC is amplified in BRCA1-associated breast cancers. Clin Cancer Res 10: 499–507.
[43]
Olaussen KA, Dunant A, Fouret P, Brambilla E, Andre F, et al. (2006) DNA repair by ERCC1 in non-small-cell lung cancer and cisplatin-based adjuvant chemotherapy. N Engl J Med 355: 983–991.
[44]
Metzger R, Leichman CG, Danenberg KD, Danenberg PV, Lenz HJ, et al. (1998) ERCC1 mRNA levels complement thymidylate synthase mRNA levels in predicting response and survival for gastric cancer patients receiving combination cisplatin and fluorouracil chemotherapy. J Clin Oncol 16: 309–316.
[45]
Niedernhofer LJ, Bhagwat N, Wood RD (2007) ERCC1 and non-small-cell lung cancer. N Engl J Med 356: 2538–2540; author reply 2540–2531.
[46]
Wachters FM, Wong LS, Timens W, Kampinga HH, Groen HJ (2005) ERCC1, hRad51, and BRCA1 protein expression in relation to tumour response and survival of stage III/IV NSCLC patients treated with chemotherapy. Lung Cancer 50: 211–219.
[47]
Cobo M, Isla D, Massuti B, Montes A, Sanchez JM, et al. (2007) Customizing cisplatin based on quantitative excision repair cross-complementing 1 mRNA expression: a phase III trial in non-small-cell lung cancer. J Clin Oncol 25: 2747–2754.
