Comparative Expression of a Set of Genes to an Internal Housekeeping Control in CDNA Amplified and not Amplified by PolyAPCR in Non-Hodgkin's Lymphoma Samples Obtained From Fine-Needle Aspiration Cytology

Comparative Expression of a Set of Genes to an Internal Housekeeping Control in CDNA Amplified and not Amplified by PolyAPCR in Non-Hodgkin's Lymphoma Samples Obtained From Fine-Needle Aspiration Cytology

Author Corbi, Fernanda Cristina Autor UNIFESP Google Scholar
Inaoka, Riguel Jun Autor UNIFESP Google Scholar
Felix, Roberta Spetic Autor UNIFESP Google Scholar
Andrade, Valeria C. C. Autor UNIFESP Google Scholar
Etto, Leina Yukari Autor UNIFESP Google Scholar
Vettore, Andre L. Autor UNIFESP Google Scholar
Franco, Marcello F. Autor UNIFESP Google Scholar
Colleoni, Gisele W. B. Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Abstract Aim: We aimed to evaluate the amount and quality of the RNA obtained from lymph nodes of non-Hodgkin lymphomas (NHLs) patients using fine-needle aspiration cytology (FNAC), and to develop strategies to overcome eventual technical drawbacks.Materials and Methods: Twenty-six patients with NHL and 10 tonsils from children submitted to tonsillectomy underwent FNAC. the aspirates were performed using both cytoaspirator (sample A) and syringe and needle (sample B). the RNA was extracted using Trizol reagent and transcribed with the Superscript kit (Invitrogen). the quality of RNA was verified through the amplification of a beta-actin 155-bp fragment.Results: Fifty-two NHL and 20 tonsil samples were analyzed. the total amount of RNA in the tonsil samples varied from <1.0 to 6.2 mu g with cytoaspirator (A) and from <1.0 to 4.7 mu g with syringe and needle (B). the total amount of RNA obtained from NHL varied from <1.0 to 6.5 mu g with cytoaspirator (A) and <1.0 to 5.5 mu g with syringe and needle. in an attempt to increase the amounts of RNA in each sample, we standardized the polyAPCR technique, which increased by 10 times the amount of cDNA in most of the test and control samples. the efficiency of the reaction was verified through the amplification of beta-actin, in which 100% of the test and control samples were amplified. When polyAPCR cDNA and nonamplified cDNA samples were paired to be evaluated by real-time PCR, using glyceraldehyde-3-phosphate dehydrogenase as the constitutive gene and nuclear factor-kappa B and NF kappa BIA as target genes, there was equivalence in the amplifications of 100% of the 15 evaluated samples.Conclusions: Our results showed that FNAC, obtained either by cytoaspirator or syringe and needle, is a good source of small amounts of RNA. the polyAPCR technique significantly increased the amount of genomic material, which might be a cDNA source for future gene expression studies.
Keywords Non-Hodgkin lymphoma
fine-needle aspiration cytology
gene expression
Language English
Sponsor Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Date 2010-03-01
Published in Diagnostic Molecular Pathology. Philadelphia: Lippincott Williams & Wilkins, v. 19, n. 1, p. 40-44, 2010.
ISSN 1052-9551 (Sherpa/Romeo, impact factor)
Publisher Lippincott Williams & Wilkins
Extent 40-44
Origin http://dx.doi.org/10.1097/PDM.0b013e3181b0b618
Access rights Closed access
Type Article
Web of Science ID WOS:000275523700007
URI http://repositorio.unifesp.br/handle/11600/32367

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