ISSN: 7027-2221

Keywords : PCR


Genetic study of different genes in the formation and metabolism of neurosteroids and cholesterol

Jenan M. Mohsen Al Mosawi; Alaa Abdul Razzaq Al Nuaimi; Farah Riadh Zayny

karbala journal of pharmaceutical sciences, Volume 7, Issue 11, Pages 44-55

Some of the neurodegenerative diseases is Parkinson's disease and Alzheimer's
disease, which can arise due to damage to nerve cells in the elderly. This study
examines the different genes that express enzymes for the formation and metabolism
of neurosteroids and cholesterol. The purpose of the study is to investigate which
genes are expressed in the CNS, and that could affect the functions of the brain via the
formation and metabolism of neurosteroids. The study used the human cell line SHSY5
as the model for neuronal cells. By knowing which genes nerve cells express
would in future be able to study the effect of different drugs can have in the formation
of steroid metabolites that affect brain functions.
Methods used in this study, RT-PCR (reverse transcription polymerase chain reaction)
and gel electrophoresis to qualitatively examine gene expression in neuroblastoma.
Results from the study suggested that SH-SY5Y cells express some but not all genes
involved in the formation and metabolism of steroid hormones and cholesterol. PCR
experiments, showed expression of the genes for the enzymes CYP19A1 and 17-
hydroxysteroid dehydrogenase (17β-HSD) and the estrogen receptor ER-β.
One conclusion that can be drawn from the results of this study is to neuroblastoma
cell line SH-SY5Y may constitute a possible cell model for future research on
hormonal effects and the effect of various drugs on the formation of neurosteroids that
influence brain functions.
The purpose of this study is to investigate whether the SH-SY5Y cells (neuroblastoma
cells) can express genes required for the formation and metabolism of cholesterol and
steroid hormones (neurosteroids). The study also intends to investigate which genes
can participate and which is expressed in the formation of steroid metabolites that
affect brain functions.

The quantification of human factor IX gene expression in transfected mammalian liver cells, employing the real time qPCR and the ELISA techniques

Zaid Al-Obaidi

karbala journal of pharmaceutical sciences, Volume 5, Issue 7, Pages 218-226

Real-time qPCR and ELISA are very sensitive powerful techniques that were commonly and universally employed in the quantification of gene expression. The aim of this study has two parts; the first part is to quantify the level of the hFIX gene expression employing the real time qPCR for quantifying the newly transcribed mRNA. The second part is to detect the translated rFIX protein using the ELISA technique. In this study the mRNA levels present in mammalian liver cells (HepG2) that were transfected with hFIX gene has been successfully quantified by qPCR using hACTB as a housekeeping gene. Also the level of the translated rFIX protein has been observed utilizing the ELISA instrument. ∆Ct and ∆∆Ct for the transfected cells were -1 and -21 respectively, while the concentration of the rFIX in these cells was 5,495 ng/ml, which is more than 800 times increase when compared with the control cells. The validation of such methods, which are used for quantification of both the gene and the translated protein, is of significant importance. Furthermore the examination of the functionality of the target protein is even more important due to the challenges associated with these techniques.