Thymoquinone regulates gene expression levels in morphine addiction pathways in opioid receptor expressing cells (U87 MG)
Background: New drugs are continuously being developed for the treatment of opioid dependence individuals. Thymoquinone is one of the drugs that exhibits inhibitory characteristics based on in vivo and in vitro models. Aim: This study further investigates the effects of thymoquinone on human g...
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Main Authors: | , , , |
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Format: | Article |
Language: | English |
Published: |
2017
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Subjects: | |
Online Access: | http://eprints.unisza.edu.my/5564/1/FH02-FP-18-12200.pdf http://eprints.unisza.edu.my/5564/ |
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Summary: | Background: New drugs are continuously being
developed for the treatment of opioid dependence
individuals. Thymoquinone is one of the drugs that
exhibits inhibitory characteristics based on in vivo
and in vitro models.
Aim: This study further investigates the effects of
thymoquinone on human global gene expression
from opioid receptor expressing cell line (OREC)
using RNA sequencing technology. The cell line that
we used was human monoglioma cancer cells (U87
MG).
Method: The quantification of RNA samples was
carried out using an Agilent 2100 Bioanalyser to
determine the RNA integrity number (RIN). Samples
with RIN>9.4 were used for further analysis. The
universal human reference RNA was used as the
common reference. The samples were treated with
morphine alone (35 µM), morphine with methadone
(162 µM), morphine with methadone and TQ (61
μM) and morphine with TQ for 48 h. The control cells
were treated with an equivalent volume of vehicle
in growth media. Total RNA was isolated from the
U87 MG cells using the RNA mini kit’s protocol
(Geneaid) and quantified using a BioDrop 2000c
spectrophotometer. After mRNA enrichment and
cDNA synthesis, the amplified cDNA fragments were
paired-end sequencing using Illumina sequencing
system with 2*150 sequencing method and subjected
to subsequent bioinformatics analysis.
Findings: The results showed that thymoquinone
upregulated phosphodiesterase 1 A (PDE1A)
genes, gamma-aminobutyric acid type A receptor
theta subunit (GABRQ) and G protein subunit beta
3 (GNG3) genes in which these genes were down
regulated by the chronic morphine.
Conclusion: These findings indicate that
thymquinone targets specific genes in morphine
addiction pathways. |
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