pro-apoptotic effects of saffron's bioactive molecules: in silico...
Post on 07-Jun-2020
5 Views
Preview:
TRANSCRIPT
Results and Discussion
Acknowledgments
Authors are grateful for Dr. Khaled Amiri, chairman of Biology Department, for
sponsoring the student’s attendance to the 5th ICDDT at Dubai.
Figure 4
Pro-apoptotic Effects of Saffron's Bioactive Molecules: In silico Analyses
Anwaar Al-Nuaimi¹, Nazar Zaki², Kamal Khazanehdari³,
Ruqaiyah A. Ali¹, Amr Amin¹
¹Biology Department, UAE University; ²Faculty of Information Technology, UAE University; ³Central Veterinary Research Laboratory, Molecular Biology & Genetics
Many studies have reported of natural antioxidants for use in food or medical materials to
replace synthetic formulations, which are restricted because of their side effects.
Natural antioxidants, found in various plants, can protect cells against oxidative damage
and may also provide an exciting preventive and therapeutic prospect for degenerative
diseases. Saffron (Fig. 1) is a spice that has been long known for its antioxidant properties
and as an anti-cancer agents for different types of cancers. The Antioxidant property of
saffron could be credited to its active ingredients (such as safranal, crocin, crocetin, and
carotene).
Apoptosis is recognized as an important mechanism in liver diseases and its down
regulation is common in cancer development including HCC. Thus, the inhibition of
apoptosis holds promise as potential therapeutic strategy against HCC.
The present study provides more details in the role of SBI on regulation of gene
expression in an animal model of liver cancer, by using RT² PCR array system (Table 1
and Fig. 2). This system is suitable for different applications including drug toxicology
studies, tumor metastasis & cancer biomarker research, as well as cytokine profiling and
inflammatory response studies. The array system allowed us through utilizing a real-time
PCR to easily examine the changes in gene expression between SBI-treated and control
samples and to quickly identify genes with significant up- or down-regulation in response
to tested SBI (Fig. 3).
The present results were consistent with our earlier study that reported a potent pro-
apoptotic effects of the crude extract of saffron both in vivo and in vitro (Fig. 4; Amin et
al., 2011).
Abstract
Alternative cancer treatment has grown into a powerful research topic for many
scientists; one of these alternative treatments is utilizing plant extracts as anticancer
remedies. Saffron is a naturally derived plant product from the dried stigma of the
Crocus sativus flower (family Iridaceae) that has significant anticancer effects that have
been reported against different cancer types. To study the mechanism of anticancer
effect of saffron, we have used gene expression analysis on number of genes in the
hepatocellular carcinoma (HCC)-induced groups. HCC is the fifth most common cancer
and the third leading cause of cancer mortality in the world. This study sheds light on
how saffron regulates rat apoptotic genes, using RT² Profiler PCR system. This array
System is the most reliable and accurate tool for analyzing the change in the level of
expression of a focused panel of genes using SYBR Green-based real-time PCR.
Introduction
Alternative cancer treatment has grown into a powerful movement that is catching on
quickly by many scientists; one of these alternative treatments is utilizing plant
extracts as anticancer remedies. Saffron is a common spice with significant anticancer
effects that has been reported against different cancer types. This study sheds light on
how saffron regulates rat apoptotic genes, using RT² Profiler PCR system. Extensive
analysis has been carried out to all apoptotic genes through the clustergram overview
which categorized the experimental groups to 6 categories into sequence of colors.
Levels of gene expressions, as represented by different colors, clearly showed that
tested saffron-based bioactive ingredients (SBI) have clear effects on number of
genes in the hepatocellular carcinoma-induced groups. SBI showed a significant
effect on Caspase-3. Caspase-3 is one of the most important genes involved in
apoptotic pathway, and thus a common target in cancer treatments. SBI effects on
levels of caspase 3 was consistent with our earlier study using saffron crude extract
which indicate that SBI tested here might play a big role in saffron’s overall effects
protecting against hepatocellular carcinoma.
Extensive analysis has been carried out to all apoptotic genes through the clustergram
overview which categorized the experimental groups to 6 categories into sequence of
colors. Levels of gene expressions, as represented by different colors, clearly showed
that tested saffron-based bioactive ingredients (SBI) have clear effects on number of
genes in the HCC-induced groups. SBI showed a significant effect on Caspase-3.
Caspase-3 is one of the most important genes involved in apoptotic pathway, and thus a
common target in cancer treatments. SBI effects on levels of caspase 3 was consistent
with our earlier study using saffron crude extract which indicate that SBI tested here
might play a big role in saffron’s overall effects protecting against HCC.
Table 1. list of genes information
associated with almost all types of
cancers and liver cancer. Genes
information from SABiosciences
database, a QIAGEN company“, all of
those genes were found to be highly
associated with apoptosis through
literature studies, and 3 of these genes
(Bcl2a1d, Tp53, casp3) have been
affected.
Fig. 3. Clutergram overview of
selected genes with their experimental groups
as follows.
Control Group Group 1 (Rats with HCC)
Group 4 (HCC + SBI-1)
Group 3 (SBI-2 alone)
Group 2 (SBI-1 alone)
Group 5 (HCC + SBI-2)
Fig. 4. (D) Lysates prepared from HepG2 cells treated with saffron for 6, 24, and 48 hours were analyzed by anti-
caspase-3, anti-IjB, anti-TNFR1, anti-pH2AX, and anti-GAPDH western blotting. GAPDH served as an internal
control for equal loading. (E) Apoptosis measurement after saffron treatment. Annexin-PI measurements of
untreated cells (control) and HepG2 cells treated with 6 mg/mL saffron for 6, 24, and 48 hours the profile
represents annexin V–fluorescein isothiocyanate (FITC) staining in the x axis and PI in the y axis.
Materials and Methods
Animals
Male wistar rats (120-200 gm) were used for this study. Experimental design.
RT² Profiler PCR Array System
Fig. 2. (a) Data sheet of
experimental groups, with all
the expressions of genes
calculated mathematically
using RT² Profiler PCR Array
system, (b) Transformation
of the data sheet to a better
overview (clustergram) using
RT² profiler PCR array analysis.
(a) (b)
References
Amin A., Hamza A., Bajbouj K., Ashraf A., Daoud S. (2011). Saffron: A Potential
Candidate for a Novel Anticancer Drug Against Hepatocellular Carcinoma. Hepatology
54: 857-867 .
Arikawa E., Prabhakar S., Zhang H., You M., Wang Y., Quellhorst G., Zeng X.,
Hung J., Yang J. Pathway-Focused Gene Expression Profiling in Toxicology, Oncology,
and Immunology Research, RT² ProfilerTM PCR Array Application Examples.
Porter A.G., Jänicke R.U. Emerging roles of caspase-3 in apoptosis. (1999). Cell death
and differentiation 6(2): 99-104.
Phil-Sun O., Sei-Jung L., Kye-Taek L. (2007). Inhibitory effect of glycoprotein isolated
from Ulmus davidiana Nakai on caspase 3 activity in 12-O-tetradecanoylphorbol 13-
acetate–treated liver cells through the reduction of intracellular reactive oxygen species.
Nutrition Research 27: 432-439.
Fig. 1. Saffron
top related