TY - JOUR
T1 - Validation and selection of new reference genes for rt-qpcr analysis in pediatric glioma of different grades
AU - Hernández-Ochoa, Beatriz
AU - Fernández-Rosario, Fabiola
AU - Castillo-Rodríguez, Rosa Angelica
AU - Marhx-Bracho, Alfonso
AU - Cárdenas-Rodríguez, Noemí
AU - Martínez-Rosas, Víctor
AU - Morales-Luna, Laura
AU - González-Valdez, Abigail
AU - Calderón-Jaimes, Ernesto
AU - Pérez de la Cruz, Verónica
AU - Rivera-Gutiérrez, Sandra
AU - Meza-Toledo, Sergio
AU - Wong-Baeza, Carlos
AU - Baeza-Ramírez, Isabel
AU - Gómez-Manzo, Saúl
N1 - Publisher Copyright:
© 2021 by the author.
PY - 2021/9
Y1 - 2021/9
N2 - Gliomas are heterogeneous, solid, and intracranial tumors that originate from glial cells. Malignant cells from the tumor undergo metabolic alterations to obtain the energy required for proliferation and the invasion of the cerebral parenchyma. The alterations in the expression of the genes related to the metabolic pathways can be detected in biopsies of gliomas of different CNS WHO grades. In this study, we evaluated the expression of 16 candidate reference genes in the HMC3 microglia cell line. Then, statistical algorithms such as BestKeeper, the comparative ΔCT method, geNorm, NormFinder, and RefFinder were applied to obtain the genes most suitable to be considered as references for measuring the levels of expression in glioma samples. The results show that PKM and TPI1 are two novel genes suitable for genic expression studies on gliomas. Finally, we analyzed the expression of genes involved in metabolic pathways in clinical samples of brain gliomas of different CNS WHO grades. RT-qPCR analysis showed that in CNS WHO grade 3 and 4 gliomas, the expression levels of HK1, PFKM, GAPDH, G6PD, PGD1, IDH1, FASN, ACACA, and ELOVL2 were higher than those of CNS WHO grade 1 and 2 glioma biopsies. Hence, our results suggest that reference genes from metabolic pathways have different expression profiles depending on the stratification of gliomas and constitute a potential model for studying the development of this type of tumor and the search for molecular targets to treat gliomas.
AB - Gliomas are heterogeneous, solid, and intracranial tumors that originate from glial cells. Malignant cells from the tumor undergo metabolic alterations to obtain the energy required for proliferation and the invasion of the cerebral parenchyma. The alterations in the expression of the genes related to the metabolic pathways can be detected in biopsies of gliomas of different CNS WHO grades. In this study, we evaluated the expression of 16 candidate reference genes in the HMC3 microglia cell line. Then, statistical algorithms such as BestKeeper, the comparative ΔCT method, geNorm, NormFinder, and RefFinder were applied to obtain the genes most suitable to be considered as references for measuring the levels of expression in glioma samples. The results show that PKM and TPI1 are two novel genes suitable for genic expression studies on gliomas. Finally, we analyzed the expression of genes involved in metabolic pathways in clinical samples of brain gliomas of different CNS WHO grades. RT-qPCR analysis showed that in CNS WHO grade 3 and 4 gliomas, the expression levels of HK1, PFKM, GAPDH, G6PD, PGD1, IDH1, FASN, ACACA, and ELOVL2 were higher than those of CNS WHO grade 1 and 2 glioma biopsies. Hence, our results suggest that reference genes from metabolic pathways have different expression profiles depending on the stratification of gliomas and constitute a potential model for studying the development of this type of tumor and the search for molecular targets to treat gliomas.
KW - Gene stability
KW - Gliomas
KW - Real-time quantitative PCR
KW - References genes
UR - http://www.scopus.com/inward/record.url?scp=85114342532&partnerID=8YFLogxK
U2 - 10.3390/genes12091335
DO - 10.3390/genes12091335
M3 - Artículo
C2 - 34573317
AN - SCOPUS:85114342532
SN - 2073-4425
VL - 12
JO - Genes
JF - Genes
IS - 9
M1 - 1335
ER -