We studied the morphological changes of the dendritic length of the pyramidal neurons of the prefrontal cortex (PFC) induced by the effect of chronic administration of caffeine in the neonatal rat. The caffeine (50 mg/kg, s.c.) was injected from day 1 after birth (P1) to day 12 (P12). The morphology of the pyramidal neurons of layer 3 of the PFC was investigated in these animals at two different ages, before puberty (P35) and after puberty (P70). Before the animals were sacrificed by using overdoses of sodium pentobarbital and being perfused intracardially with 0.9% saline, the locomotor activity in a novel environment was measured. The brains were then removed, processed by the Golgi-Cox stain, and analyzed by the Sholl method. The dendritic morphology clearly showed that the neonatal animals administered caffeine showed an increase in the dendritic length of the pyramidal neurons of the PFC when compared with the control animals at both ages. The present results suggest that neonatal administration of caffeine may in part affect the dendritic morphology of the pyramidal cells of this limbic structure and this effect persists after puberty and may be implicated in several brain processes.

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.

BackgroundStudies of alternative mRNA splicing (AS) in health and disease have yet to yield the complete picture of protein diversity and its role in physiology and pathology. Some forms of cancer appear to be associated to certain alternative mRNA splice variants, but their role in the cancer development and outcome is unclear.MethodsWe examined AS profiles by means of whole genome exon expression microarrays (Affymetrix GeneChip 1.0) in ovarian tumors and ovarian cancer-derived cell lines, compared to healthy ovarian tissue. Alternatively spliced genes expressed predominantly in ovarian tumors and cell lines were confirmed by RT-PCR.ResultsAmong several significantly overexpressed AS genes in malignant ovarian tumors and ovarian cancer cell lines, the most significant one was that of the zinc finger protein ZNF695, with two previously unknown mRNA splice variants identified in ovarian tumors and cell lines. The identity of ZNF695 AS variants was confirmed by cloning and sequencing of the amplicons obtained from ovarian cancer tissue and cell lines.ConclusionsAlternative ZNF695 mRNA splicing could be a marker of ovarian cancer with possible implications on its pathogenesis.

Low levels of oxygen (hypoxia) have been reported in solid tumours. This hypoxic microenvironment modulates the expression of genes linked to a more aggressive disease. However, it is unclear if the expression of drug‐metabolizing enzymes as cytochromes P450 (CYPs) is affected by hypoxia in cancer. We aimed to define which cytochromes are affected by hypoxia using a liver cancer model in vitro. For this purpose, we assessed whole‐genome expression microarrays of HepG2 liver cancer cell line from free repository databases, looking for gene expression hypoxia‐associated profiles and selected those cytochromes with significant differences. Then, we corroborated their mRNA expression and protein levels by RT‐qPCR and western blot, respectively, as well as immunofluorescence. Based on microarray analysis, we found that the expression of CYP2S1 and CYP24A1 were up‐regulated with at least twice fold change compared with normoxia. The levels of mRNA and protein of CYP2S1 and CYP24A1 were increased significantly in hypoxic conditions (P < .05), and this tendency was also observed by immunofluorescence assays. Our data show that the expression of cytochromes CYP2S1 and CYP24A1 are induced in hypoxia, being the first time that CYP24A1 expression is associated with tumour hypoxia; which might have consequences in cancer progression and drug resistance. Significance of the study Hypoxia is among the most important factors for cellular adaptation to stress. Especially in cancer, a major public health issue, hypoxia plays a substantial role in angiogenesis, metastasis and resistance to therapy. Tumoral hypoxia has been described at least in the brain, breast, cervical, liver, renal, lung, pancreatic and renal cancer. However, the understanding of how hypoxia drives cancer progression is still a major challenge. One emerging question is the role of hypoxia over the expression of drug‐metabolizing enzymes, with a significant impact on drug treatment. In this context, our paper focus on the effect of hypoxia on CYPs, which is an essential group of drug‐metabolizing enzymes. We show that hypoxia induces the expression of two members of the CYPs family: CYP2S1 and CYP24A1. Importantly, CYP2S1 is a major metabolizer of carcinogenic substances being relevant that hypoxia could promote this function. Interestingly, CYP24A1 limits the action of the active form of vitamin D, which is an anti‐proliferative factor in cancer. Our evidence shows for the first time that hypoxia can induce CYP24A1 expression, with a potential effect on cancer progression. Our contribution clarifies a particular effect of tumoral hypoxia and the implications will be useful in the understanding of the progression of cancer, the resistance to treatment and the development of alternative therapies.

Background B‐cell acute lymphoblastic leukemia (B‐ALL) is the most commonly diagnosed childhood malignancy worldwide and is especially common in Mexico. Additionally, the number of cases has increased in recent years. Thus, it is very important to develop molecular strategies to diagnose leukemia. The aim of this study was to investigate MYB expression and to determine its impact on the diagnosis of B‐ALL. Methods We analyzed the B‐ALL gene expression profile by microarray data mining. Bioinformatics analysis was performed to identify the genes that are overexpressed in leukemia. We determined that MYB was highly expressed in leukemia. Then, we validated MYB expression in 70 patients with B‐ALL and in 16 healthy controls (HCs) using qRT‐PCR. The results were statistically analyzed using the Kolmogorov‐Smirnov Z test, Mann‐Whitney U test, receiver operating characteristic curves, and the Youden index. Results The microarrays showed that MYB was overexpressed in B‐ALL patients with a fold change of 57.8728 and a P value of 2.56−195. MYB expression showed great variability among the patients analyzed. However, compared to the HCs, the B‐ALL patients had a P value < .0001, an area under the curve of 0.813, and a Youden index of 1.46, indicating the statistical significance. Conclusion MYB expression in B‐ALL cells could be a potential molecular marker for childhood leukemia.