Increased Expression of Thymidylate Synthetase (TS), Ubiquitin Specific Protease 10 (USP10) and Survivin is Associated with Poor Survival in Glioblastoma Multiforme (GBM)
The limited success of empirically designed treatment paradigms for patients diagnosed with glioblastoma multiforme (GBM) emphasizes the need for rationally designed treatment strategies based on the molecular profile of tumor samples and their correlation to clinical parameters.
In the current study, we utilize a novel real-time quantitative low density array (RTQ-LDA) to identify differentially expressed genes in de novo GBM tissues obtained from patients with distinctly different clinical outcomes. Total RNA was isolated from a cohort of 21 GBM specimens obtained from patients with either good (long-term survival (LTS) >36 months post surgery, n = 8) or poor (died of the disease (DOD) <24 months post surgery, n = 13) prognosis. Non-neoplastic brain tissue (n = 5) was obtained from patients who underwent surgery for refractory epilepsy. Demographic data was assessed for correlation with survival using Cox proportional hazards models. Sufficient RNA was available to use RTQ-LDA to quantify the expression of 93 independent genes in 5␣LTS, 4 DOD, and 5 non-neoplastic brain samples. The eight differentially expressed genes identified by RTQ-LDA in LTS versus DOD (P ≤ 0.050) were subsequently quantified in all 21 GBM samples by real-time quantitative PCR (RTQ).
A correlation between younger patients and good prognosis was demonstrated (P ≤ 0.05). The combination of RTQ-LDA and RTQ identified thymidylate synthetase (TS), ubiquitin specific protease 10 (USP10), and survivin as significantly over-expressed (P ≤ 0.050) in DOD compared to LTS patients. Ribonucleotide reductase subunit M2 (RRM2) was identified as tumor-specific, but not associated with survival.
Taken collectively, TS, USP10, survivin and RRM2 may be useful as prognostic indicators and/or in the development of rationally designed treatment protocols.
KeywordsGlioblastoma multiforme Glioma Low density array Real-time quantitative PCR Ribonucleotide reductase subunit M2 Survival Survivin Thymidylate synthetase Ubiquitin specific protease 10
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We thank Cecil R. Stockard for his excellent immunohistochemical assistance. Supported by American Cancer Society RSG-04-030-01-CCE and NCI P50 CA 097247.
- 3.Fazeny-Dorner B, Gyries A, Rossler K, Ungersbock K, Czech T, Budinsky A, Killer M, Dieckmann K, Piribauer M, Baumgartner G, Prayer D, Veitl M, Muhm M, Marosi C (2003) Survival improvement in patients with glioblastoma multiforme during the last 20 years in a single tertiary-care center. Wien Klin Wochenschr 115(11):389–397PubMedCrossRefGoogle Scholar
- 7.van den Boom J, Wolter M, Kuick R, Misek DE, Youkilis AS, Wechsler DS, Sommer C, Reifenberger G, Hanash SM (2003) Characterization of gene expression profiles associated with glioma progression using oligonucleotide-based microarray analysis and real-time reverse transcription-polymerase chain reaction. Am J Pathol 163(3):1033–1043PubMedGoogle Scholar
- 8.Nutt CL, Mani DR, Betensky RA, Tamayo P, Cairncross JG, Ladd C, Pohl U, Hartmann C, McLaughlin ME, Batchelor TT, Black PM, von Deimling A, Pomeroy SL, Golub TR, Louis DN (2003) Gene expression-based classification of malignant gliomas correlates better with survival than histological classification. Cancer Res 63(7):1602–1607PubMedGoogle Scholar
- 9.Fuller GN, Hess KR, Rhee CH, Yung WK, Sawaya RA, Bruner JM, Zhang W (2002) Molecular classification of human diffuse gliomas by multidimensional scaling analysis of gene expression profiles parallels morphology-based classification, correlates with survival, and reveals clinically-relevant novel glioma subsets. Brain Pathol 12(1):108–116PubMedCrossRefGoogle Scholar
- 11.Burton EC, Lamborn KR, Forsyth P, Scott J, O’Campo J, Uyehara-Lock J, Prados M, Berger M, Passe S, Uhm J, O’Neill BP, Jenkins RB, Aldape KD (2002) Aberrant p53, mdm2, and proliferation differ in glioblastomas from long-term compared with typical survivors. Clin Cancer Res 8(1):180–187PubMedGoogle Scholar
- 12.Davis FG, Freels S, Grutsch J, Barlas S, Brem S (1998) Survival rates in patients with primary malignant brain tumors stratified by patient age and tumor histological type: an analysis based on Surveillance, Epidemiology, and End Results (SEER) data, 1973–1991. J Neurosurg 88(1):1–10PubMedGoogle Scholar
- 19.Steg A, Wang W, Blanquicett C, Grunda JM, Eltoum IA, Wang K, Buchsbaum DJ, Vickers SM, Russo S, Diasio RB, Frost AR, LoBuglio AF, Grizzle WE, Johnson MR (2006) Multiple gene expression analyses in paraffin-embedded tissues by TaqMan low-density array: application to hedgehog and Wnt pathway analysis in ovarian endometrioid adenocarcinoma. J Mol Diagn 8(1):76–83PubMedGoogle Scholar
- 22.Zar JH (1999) Biostatistical analysis, 4th ed. Prentice-Hall, Upper Saddle RiverGoogle Scholar
- 23.Allison DB, Gadbury G, Heo M, Fernandez J, Lee C, Prolla TA, Weindruch R (2002) A mixture model approach for the analysis of microarray gene expression data. Comput Stat Data Anal (39):1–20Google Scholar
- 25.Klecka WR (1980) Discriminant analysis. Sage University Paper Series on Quantitative Applications in the Social Sciences Series No. 07-019Google Scholar
- 26.Fisher RA (1936) The use of multiple measurements in taxonomic problems. Ann Eugenic (7):179–188Google Scholar
- 33.Fulda S, Debatin KM (2004) Sensitization for anticancer drug-induced apoptosis by the chemopreventive agent resveratrol. Oncogene 23(40):6702–6711Google Scholar
- 40.Menei P, Capelle L, Guyotat J, Fuentes S, Assaker R, Bataille B, Francois P, Dorwling-Carter D, Paquis P, Bauchet L, Parker F, Sabatier J, Faisant N, Benoit JP (2005) Local and sustained delivery of 5-fluorouracil from biodegradable microspheres for the radiosensitization of malignant glioma: a randomized phase II trial. Neurosurgery 56(2):242–248 (discussion 242–248)PubMedCrossRefGoogle Scholar
- 42.Blanquicett C, Buchsbaum DJ, Saif MW, Eloubeidi M, Vickers SM, Chhieng DC, Carpenter MD, Sellers JC, Russo S, Diasio RB, Johnson MR (2005) Antitumor efficacy of capecitabine and celecoxib in irradiated and lead shielded contralateral human BxPC-3 pancreatic cancer xenografts: clinical implications of abscopal effects in a metastatic model. Clin Cancer Res (submitted)Google Scholar
- 43.Blanquicett C, Gillespie GY, Nabors LB, Miller CR, Bharara S, Buchsbaum DJ, Diasio RB, Johnson MR (2002) Induction of thymidine phosphorylase in both irradiated and shielded, contralateral human U87MG glioma xenografts: implications for a dual modality treatment using capecitabine and irradiation. Mol Cancer Ther 1(12):1139–1145PubMedGoogle Scholar
- 44.Newman AJ, Fiveash J, Rosenfeld S, Johnson M, Diasio R, Wang W, Cockrell-Donohue A, Nabors LB (2004) A phase I study of capecitabine, concurrent radiotherapy (RT) for patients with newly diagnosed glioblastoma multiforme (GBM). In: Grunberg SM, Gary A, Whippen D (eds) Proc Am Soc Clin Oncol, New Orleans LA, June 5–8 2004. Lisa Greaves, American Society of Clinical Oncology, Alexandria, 2004, 116 ppGoogle Scholar
- 55.Lee Y, Vassilakos A, Feng N, Lam V, Xie H, Wang M, Jin H, Xiong K, Liu C, Wright J, Young A (2003) GTI-2040, an antisense agent targeting the small subunit component (R2) of human ribonucleotide reductase, shows potent antitumor activity against a variety of tumors. Cancer Res 63(11):2802–2811PubMedGoogle Scholar
- 58.Stupp R, Mason WP, Van Den Bent MJ, Weller M, Fisher B, Taphoorn M, Brandes AA, Cairncross JG, Lacombe D, Mirimanoff RO (2004) Concomitant and adjuvant temozolomide (TMZ) and radiotherapy (RT) for newly diagnosed glioblastoma multiforme (GBM). Conclusive results of a randomized phase III trail by the EORTC Brain & RT Groups and NCIC Clinical Trials Group. In: Grunberg SM, Gary A, Whippen D (eds) Proc Am Soc Clin Oncol, New Orleans, Louisiana, June 5–8 2004. Lisa Greaves, American Society of Clinical Oncology, Alexandria, 2004, p 1Google Scholar