Effective Mixed-Type Tissue Crusher and Simultaneous Isolation of RNA, DNA, and Protein from Solid Tissues Using a TRIzol-Based Method

Kelly Karoline dos Santos, Isabelle Watanabe Daniel, Letícia Carani Delabio, Manoella Abrão da Costa, Júlia de Paula Dutra, Bruna Estelita Ruginsk, Jeanine Marie Nardin, Louryana Padilha Campos, Fabiane Gomes de Moraes Rego, Geraldo Picheth, Glaucio Valdameri and Vivian Rotuno Moure ()
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Kelly Karoline dos Santos: Graduate Program in Pharmaceutical Sciences, Laboratory of Cancer Drug Resistance, Federal University of Parana, Curitiba CEP 80210-170, PR, Brazil
Isabelle Watanabe Daniel: Graduate Program in Pharmaceutical Sciences, Laboratory of Cancer Drug Resistance, Federal University of Parana, Curitiba CEP 80210-170, PR, Brazil
Letícia Carani Delabio: Graduate Program in Pharmaceutical Sciences, Laboratory of Cancer Drug Resistance, Federal University of Parana, Curitiba CEP 80210-170, PR, Brazil
Manoella Abrão da Costa: Graduate Program in Pharmaceutical Sciences, Laboratory of Cancer Drug Resistance, Federal University of Parana, Curitiba CEP 80210-170, PR, Brazil
Júlia de Paula Dutra: Graduate Program in Pharmaceutical Sciences, Laboratory of Cancer Drug Resistance, Federal University of Parana, Curitiba CEP 80210-170, PR, Brazil
Bruna Estelita Ruginsk: Graduate Program in Pharmaceutical Sciences, Laboratory of Cancer Drug Resistance, Federal University of Parana, Curitiba CEP 80210-170, PR, Brazil
Jeanine Marie Nardin: School of Medicine and Life Sciences, Pontifical Catholic University of Parana, Curitiba CEP 80215-901, PR, Brazil
Louryana Padilha Campos: Graduate Program in Pharmaceutical Sciences, Federal University of Parana, Curitiba CEP 80210-170, PR, Brazil
Fabiane Gomes de Moraes Rego: Graduate Program in Pharmaceutical Sciences, Federal University of Parana, Curitiba CEP 80210-170, PR, Brazil
Geraldo Picheth: Graduate Program in Pharmaceutical Sciences, Federal University of Parana, Curitiba CEP 80210-170, PR, Brazil
Glaucio Valdameri: Graduate Program in Pharmaceutical Sciences, Laboratory of Cancer Drug Resistance, Federal University of Parana, Curitiba CEP 80210-170, PR, Brazil
Vivian Rotuno Moure: Graduate Program in Pharmaceutical Sciences, Laboratory of Cancer Drug Resistance, Federal University of Parana, Curitiba CEP 80210-170, PR, Brazil

J, 2025, vol. 8, issue 1, 1-19

Abstract: One of the major challenges of studying biomarkers in tumor samples is the low quantity and quality of isolated RNA, DNA, and proteins. Additionally, the extraction methods ideally should obtain macromolecules from the same tumor biopsy, allowing better-integrated data interpretation. In this work, an in-house, low-cost, mixed-type tissue crusher combining blade and beating principles was made and the simultaneous isolation of macromolecules from human cells and tissues was achieved using TRIzol. RT-qPCR, genotyping, SDS-PAGE, and Western blot analysis were used to validate the approach. For tissue samples, RNA, DNA, and proteins resulted in an average yield of 677 ng/mg, 225 ng/mg, and 1.4 µg/mg, respectively. The same approach was validated using cell lines. The isolated macromolecule validation included the detection of mRNA levels of ATP-binding cassette (ABC) transporters through RT-qPCR, genotyping of TNFR1 (rs767455), and protein visualization through SDS-PAGE following Coomassie blue staining and Western blot. This work contributed to filling a gap in knowledge about TRIzol efficiency for the simultaneous extraction of RNA, DNA, and proteins from a single human tissue sample. A low-cost, high yield, and quality method was validated using target biomarkers of multidrug resistance mechanisms. This approach might be advantageous for future biomarker studies using different tissue specimens.

Keywords: cancer; human tissues; TRIzol; total RNA; DNA; proteins; MDR; ABC transporters (search for similar items in EconPapers)
JEL-codes: I1 I10 I12 I13 I14 I18 I19 (search for similar items in EconPapers)
Date: 2025
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