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Open AccessArticle

Identification and Validation of Novel Reference Genes in Acute Lymphoblastic Leukemia for Droplet Digital PCR

1
Experimental Oncology Laboratory, Research Department, National Institute of Pediatrics, Mexico City 04530, Mexico
2
Research Methodology Department, National Institute of Pediatrics, Mexico City 04530, Mexico
*
Authors to whom correspondence should be addressed.
Genes 2019, 10(5), 376; https://doi.org/10.3390/genes10050376
Received: 25 March 2019 / Revised: 3 May 2019 / Accepted: 7 May 2019 / Published: 17 May 2019
(This article belongs to the Section Human Genomics and Genetic Diseases)
Droplet digital PCR is the most robust method for absolute nucleic acid quantification. However, RNA is a very versatile molecule and its abundance is tissue-dependent. RNA quantification is dependent on a reference control to estimate the abundance. Additionally, in cancer, many cellular processes are deregulated which consequently affects the gene expression profiles. In this work, we performed microarray data mining of different childhood cancers and healthy controls. We selected four genes that showed no gene expression variations (PSMB6, PGGT1B, UBQLN2 and UQCR2) and four classical reference genes (ACTB, GAPDH, RPL4 and RPS18). Gene expression was validated in 40 acute lymphoblastic leukemia samples by means of droplet digital PCR. We observed that PSMB6, PGGT1B, UBQLN2 and UQCR2 were expressed ~100 times less than ACTB, GAPDH, RPL4 and RPS18. However, we observed excellent correlations among the new reference genes (p < 0.0001). We propose that PSMB6, PGGT1B, UBQLN2 and UQCR2 are housekeeping genes with low expression in childhood cancer. View Full-Text
Keywords: references genes; droplet digital PCR; gene expression; leukemia references genes; droplet digital PCR; gene expression; leukemia
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Villegas-Ruíz, V.; Olmos-Valdez, K.; Castro-López, K.A.; Saucedo-Tepanecatl, V.E.; Ramírez-Chiquito, J.C.; Pérez-López, E.I.; Medina-Vera, I.; Juárez-Méndez, S. Identification and Validation of Novel Reference Genes in Acute Lymphoblastic Leukemia for Droplet Digital PCR. Genes 2019, 10, 376.

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