Measuring Expression Levels of Small Regulatory RNA Molecules from Body Fluids and Formalin-Fixed, Paraffin-Embedded Samples

Author(s):  
Adrienn Gyongyosi ◽  
Otto Docs ◽  
Zsolt Czimmerer ◽  
Laszlo Orosz ◽  
Attila Horvath ◽  
...  
Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 5197-5197
Author(s):  
Hua Zhong

Abstract Abstract 5197 Introduction Diffuse large B cell lymphoma (DLBCL) is an aggressive malignancy that accounts for nearly 40% of all lymphoid tumors. Both miR-155 and miR-146a expression levels are suggested to be regulated by NF-KB expression in DLBCL. In present study, the expression of miR-155 and miR-146a were investigated to evaluate the prognosis in DLBCL patients. Patients and Methods The expression levels of miR-155 and miR-146a in formalin-fixed/paraffin-embedded tissue of patients with DLBCL (n=50) have been compared with reactive hyperplasia lymphoid nodes (n=20) by using real-time polymerase chain reaction. Clinical characters of patients such as LDH, β2-MG, International Prognostic Index (IPI), ECOG score and c-myc expression levels have also been investigated. Results 1) The expression levels of miR-155 and miR-146a were significantly higher in DLBCL patients than control (P<0.001). (Fig 1) 2) The expression levels of miR-155 and miR-146a were different between germinal center B-cell like DLBCL (GCB-DLBCL) and activated B-cell-like DLBCL (ABC-DLBCL), but the difference had no significance (P>0.05). 3) There was correlation between the expression of miR-155 and miR-146a in DLBCL patients (r =0.639, P<0.001). 4) There was positive correlation in the expression of miR-146a with LDH, β2-MG, IPI, ECOG score and c-myc expression levels in DLBCL patients. 5) Lower expression levels of miR-155 and miR-146a were found to be associated with high complete remission (CR) rate and overall response (OR) rate in DLBCL patients (P<0.05). 6) Lower expression levels of miR-146a in DLBCL patients were only associated with better 5 years progression free survival (5y-PFS) (P=0.044), but lower expression levels of miR-155 in DLBCL patients were associated with better 5y-PFS (P=0.013) and 5 years overall survival (5y-OS) (P=0.007). 7) Multivariate analysis using a Cox proportional hazard model confirmed that either miR-155 or miR-146a was the independent predictor. (For miR-155 5y-PFS, Hazards ratio 0.260, 95% CI, 0.058–0.801; for miR-146a 5y-PFS, Hazards ratio 0.251, 95% CI, 0.068–0.922) 8) Compared to CHOP protocol, the higher miR-155 expression level patients who chose R-CHOP protocol for treatment achieved better 3y-PFS and 3y-OS (P<0.05). Conclusions The expression levels of miR-155 and miR-146a were higher in DLBCL patients. Moreover, high expressions of these two microRNAs were associated with the disease progression. Patients with higher miR-155 expression levels may be benefit from the protocol including Rituximab. Disclosures: No relevant conflicts of interest to declare.


2015 ◽  
Vol 112 (48) ◽  
pp. 14924-14929 ◽  
Author(s):  
Pierluigi Gasparini ◽  
Luciano Cascione ◽  
Lorenza Landi ◽  
Stefania Carasi ◽  
Francesca Lovat ◽  
...  

microRNAs (miRNAs) can act as oncosuppressors or oncogenes, induce chemoresistance or chemosensitivity, and are major posttranscriptional gene regulators. Anaplastic lymphoma kinase (ALK), EGF receptor (EGFR), and V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) are major drivers of non-small cell lung cancer (NSCLC). The aim of this study was to assess the miRNA profiles of NSCLCs driven by translocated ALK, mutant EGFR, or mutant KRAS to find driver-specific diagnostic and prognostic miRNA signatures. A total of 85 formalin-fixed, paraffin-embedded samples were considered: 67 primary NSCLCs and 18 matched normal lung tissues. Of the 67 primary NSCLCs, 17 were echinoderm microtubule-associated protein-like 4–ALK translocated (ALK+) lung cancers; the remaining 50 were not (ALK−). Of the 50 ALK− primary NSCLCs, 24 were EGFR and KRAS mutation-negative (i.e., WT; triple negative); 11 were mutant EGFR (EGFR+), and 15 were mutant KRAS (KRAS+). We developed a diagnostic classifier that shows how miR-1253, miR-504, and miR-26a-5p expression levels can classify NSCLCs as ALK-translocated, mutant EGFR, or mutant KRAS versus mutation-free. We also generated a prognostic classifier based on miR-769-5p and Let-7d-5p expression levels that can predict overall survival. This classifier showed better performance than the commonly used classifiers based on mutational status. Although it has several limitations, this study shows that miRNA signatures and classifiers have great potential as powerful, cost-effective next-generation tools to improve and complement current genetic tests. Further studies of these miRNAs can help define their roles in NSCLC biology and in identifying best-performing chemotherapy regimens.


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