title: Predictive and prognostic value of TLR9 and NFKBIA gene expression as potential biomarkers for human glioma diagnosis.
abstract: BACKGROUND
  Malignant gliomas are the most common form of primary intracranial tumors with the highest mortality rates. Various gene alterations are considered as prognostic markers in glioma. But, the relevant molecular mechanisms in this setting are not well-understood.
  
  
  OBJECTIVE
  The aim of this study was to assess the association and prognostic value of TLR9 and NFKBIA with clinical significance and also their impact on patient survival in human glioma.
  
  
  METHODS
  Expression of TLR9 and NFKBIA mRNA in the tissues was determined by immunohistochemistry and qRT-PCR methods. Kaplan-Meier curves and Cox proportional hazards regression model were used to assess the association of TLR9 and NFKBIA with clinical outcomes of patients.
  
  
  RESULTS
  Quantitative real-time PCR analysis showed that TLR9 mRNAs is markedly expressed in glioma tissues than in non-neoplastic tissues (mean±SD: 3.26±0.40 vs. 0.71±0.36, P<0.001). There was also a significant difference between TLR9 mRNAs and high grade glioma (P<0.001).NFKBIA mRNAs was significantly identified in non-neoplastic tissues compared with glioma specimens (mean±SD: 2.76±0.30 vs. 0.94±0.35, P<0.001). Lower levels of NFKBIA mRNA were significantly related to advanced grade of gliomas (P<0.001). Furthermore, Immunoreactivity for high expression of TLR9 was detected in 65% of cases (26/40) that was associated with high grade glioma (P=0.001). No statistically significant correlation was found between TLR9 and other clinical parameters (P>0.05). Immunoreactivity for high expression of NFKBIA was observed in 32.5% (13/40) of cases and NFKBIA expression was decreased in patients with high grad glioma (P=0.014). There was no significant correlation between NFKBIA protein expression and age, sex, and relapse. The Kaplan-Meier analysis indicated that patients with high expression of TLR9 and low expression of NFKBIA are significantly related to poorer OS (P<0.001). In addition, the multivariate Cox regression model revealed that TLR9 and NFKBIA protein expressions (low/high) and tumor grade were potentially an independent predictor of survival in patients (hazard ratio, 2.132, 2.411, 2.13 [95% confidence interval, 1.825-3.782, 1.61-3.231, 1.542-3.92]; P=0.012,P=0.018, P=0.001).
  
  
  CONCLUSIONS
  These data indicate that TLR9 and NFKBIA protein expressions act as independent predictor of survival for the diagnosis of glioma and a prognostic biomarker for those with a tumor at an advanced pathological grade.
authors.affiliation: Faculty of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Ata
authors.initials: A
authors.lastname: Miyar
authors.affiliation: Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Iman
authors.initials: I
authors.lastname: Habibi
authors.affiliation: Students' Research Committee, Baqiyatallah University of Medical Sciences, Tehran, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Ebrahimi
authors.affiliation: Brain and Spinal Cord Injury Research Center, Neuroscience Institute, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Delaram
authors.initials: D
authors.lastname: Mansourpour
authors.affiliation: Cellular & Molecular Research Center, Kurdistan University of Medical Sciences, Sanandaj, Iran.
authors.forename: Aram
authors.initials: A
authors.lastname: Mokarizadeh
authors.affiliation: Graduated from the Faculty of Veterinary Medicine, Islamic Azad University, Karaj Branch, Karaj, Iran.
authors.forename: Alireza
authors.initials: A
authors.lastname: Rajabi
authors.affiliation: Veterinarian in Private Sector. Electronic address: rozhinfarshgar3000@gmail.com.
authors.forename: Rozhin
authors.initials: R
authors.lastname: Farshgar
authors.affiliation: Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Eshaghzadeh
authors.affiliation: Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran.
authors.forename: Mohamad
authors.initials: M
authors.lastname: Zamani-Ahmadmahmudi
authors.affiliation: Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Seyed Mohamad Hossein Tabatabaei
authors.initials: SM
authors.lastname: Nodushan
mesh_terms: D000328:Adult; D000367:Age Factors; D001932:Brain Neoplasms / Q000175:diagnosis / Q000235:genetics* / Q000601:surgery; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D005820:Genetic Testing; D005910:Glioma / Q000175:diagnosis / Q000235:genetics* / Q000601:surgery; D006801:Humans; D020672:I-kappa B Proteins / Q000235:genetics / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D000072000:NF-KappaB Inhibitor alpha; D011379:Prognosis; D016016:Proportional Hazards Models; D012333:RNA, Messenger / Q000378:metabolism; D051217:Toll-Like Receptor 9 / Q000235:genetics / Q000378:metabolism*
chemical_list: D020672:I-kappa B Proteins; C000606204:NFKBIA protein, human; D012333:RNA, Messenger; C495371:TLR9 protein, human; D051217:Toll-Like Receptor 9; D000072000:NF-KappaB Inhibitor alpha
keywords: Biomarker; Glioma; IHC; PCR; Patient; Survival; TLR9 and NFKBIA