|Validation of Total RNA Spin-column and Non-column Extraction Methods for microRNA Expression Analyses in Formalin-fixed Paraffin-Embedded Liver Samples|
|Zahra Asefy1,2,3, Alireza Abhari2, Behrooz Shokuhi1, Zeinab Latif2, Saeed Nazari Soltan Ahmad1,2, Sirus Hoseinnejhad4, Naser Samadi2, Mohammad Nouri1,2|
|1Liver and Gastrointestinal Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
2Department of Biochemistry and Clinical Laboratories, Tabriz University of Medical Sciences, Tabriz, Iran
3Student's Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran
4Department of Biochemistry, Baku State University, Baku, Azerbaijan
CJMB 2019; 6: 346-349
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Keywords : Spin column extraction, miRNA, FFPE
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Objectives: MicroRNAs are small (19–23nt) noncoding RNAs that adjust gene expression post-transcriptionally. In addition, microRNA (miRNA) profile modification is revealed in various abnormalities including cancer. Formalin-fixed paraffin-embedded (FFPE) material archives are valued for searching human diseases. Therefore, the present study aimed to investigate FFPE samples for miRNA expression, as an alternative analyte for gene expression with the growing therapeutic and diagnostic potential. Further, the validity of RNA isolation kits was evaluated in order to identify a preferred method.
Materials and Methods: The two extracted methods were qualitatively and quantitatively assessed in order to compare the archived specimens between these methods. The total RNA and gene expression were quantified in each method as well.
Results: Based on the results, the level of RNA extracted from FFPE tissues and real-time polymerase chain reaction (RT-PCR) of hsa-miR-623, hsa-miR-515-5pin miRNeasy FFPE (P value < 0.047) was significantly higher than that in RNAX Plus (P < 0.084).
Conclusions: In general, the spin column method for miRNA expression profile had better expression compared to the non-column method. The results further confirmed the validity of FFPE tissues as an appropriate resource for miRNA analyses.
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