Analysis of housekeeping genes in the peripheral blood of retinoblastoma patients

Abstract

Retinoblastoma is a pediatric neoplasia with a high incidence in non-developed countries. Nowadays the diagnosis is clinical and unfortunately in advanced stages of the diseases, which puts the child´s life risk. The search for molecular diagnosis on retinoblastoma is necessary. Due to their location and tendency to migrate, biopsies of retinoblastoma are not recommended then, peripheral blood samples can be a good source for the search of biomarker. RT qPCR is a sensitive method for gene expression quantification; in order to achieve optimal results, a crucial step is the reference gene choice that cannot be neglected under any circumstance. Six of the most commonly used housekeeping genes; GAPDH, HPRT1, B2M, TBP, RPL13a and 18S, were tested in the blood samples of patients diagnosed with retinoblastoma and healthy controls.The HPRT and TBP were found the most reliable genes whereas GAPDH, that is one of the most commonly used genes for normalisation, together with B2M, RPL13a and 18S have to be avoided. Using the selected reference genes, the Rb mRNA showed significant differences between patients and healthy children whereas no differences were found using to control groups. In the present study, we validate blood samples of patients with retinoblastoma.

Investigation of chicken housekeeping genes using next-generation sequencing data

“…In the present work, as compared to those of other tissues, greater number of WU-HKGs were relatively constant across all datasets of blood. Studying human peripheral blood, Martínez-Sánchez et al (2019) found Hprt and Tbp as the most reliable genes. They suggested the utilization of Gapdh , B2m , and Rpl13a genes to be avoided.…”

“…Frontiers in Genetics frontiersin.org datasets of blood. Studying human peripheral blood, Martínez-Sánchez et al (2019) found Hprt and Tbp as the most reliable genes. They suggested the utilization of Gapdh, B2m, and Rpl13a genes to be avoided.…”

Retinoblastoma: from discovery to clinical management

“…These findings suggest a possible hereditary component; therefore, the mRNA levels of the former genes were measured in family members and compared to the patients. Strikingly, the results indicated a higher level of the p53 mRNA in the family members than in controls [55]. Given the hypothetical diagnostic power of the previously mentioned biomarkers, it would be interesting to develop further validation and implementation in the clinic.…”

“…Loss of the RB gene can induce apoptosis, representing a mechanism against the proliferation of potentially malignant cells [53]. Cancer cells can then proliferate and overcome the RB1 loss-dependent apoptosis when accompanied by other events such as abrogation of the p53 pathways [54][55][56]. RB also has pro-apoptotic behaviours; indirectly, it was shown that RB can defeat the MDM2-anti-apoptotic function on p53-induced apoptosis in 1999.…”