Abstract: Harmalol administration caused remarkable reduction in proliferation of HepG2 cells with GI50 of 14.2 mM, without showing much cytotoxicity in embryonic liver cell line, WRL-68. Data from circular dichroism and differential scanning calorimetric analysis of harmalol-CT DNA complex shows conformational changes with prominent CD perturbation and stabilization of CT DNA by 8 oC. Binding constant and stoichiometry was also calculated using the above biophysical techniques. Further, dose dependent apoptotic induction ability of harmalol was studied in HepG2 cells using different biochemical assays. Generation of ROS, DNA damage, changes in cellular external and ultramorphology, alteration of membrane, formation of comet tail, decreased mitochondrial membrane potential and a significant increase in Sub Go/G1 population made the cancer cell, HepG2, prone to apoptosis. Up regulation of p53 and caspase 3 further indicated the apoptotic role of harmalol.
Abstract: Array-based gene expression analysis is a powerful
tool to profile expression of genes and to generate information on
therapeutic effects of new anti-cancer compounds. Anti-apoptotic
effect of thymoquinone was studied in MCF7 breast cancer cell line
using gene expression profiling with cDNA microarray. The purity
and yield of RNA samples were determined using RNeasyPlus Mini
kit. The Agilent RNA 6000 NanoLabChip kit evaluated the quantity
of the RNA samples. AffinityScript RT oligo-dT promoter primer
was used to generate cDNA strands. T7 RNA polymerase was used to
convert cDNA to cRNA. The cRNA samples and human universal
reference RNA were labelled with Cy-3-CTP and Cy-5-CTP,
respectively. Feature Extraction and GeneSpring softwares analysed
the data. The single experiment analysis revealed involvement of 64
pathways with up-regulated genes and 78 pathways with downregulated
genes. The MAPK and p38-MAPK pathways were
inhibited due to the up-regulation of PTPRR gene. The inhibition of
p38-MAPK suggested up-regulation of TGF-ß pathway. Inhibition of
p38-MAPK caused up-regulation of TP53 and down-regulation of
Bcl2 genes indicating involvement of intrinsic apoptotic pathway.
Down-regulation of CARD16 gene as an adaptor molecule regulated
CASP1 and suggested necrosis-like programmed cell death and
involvement of caspase in apoptosis. Furthermore, down-regulation
of GPCR, EGF-EGFR signalling pathways suggested reduction of
ER. Involvement of AhR pathway which control cytochrome P450
and glucuronidation pathways showed metabolism of Thymoquinone.
The findings showed differential expression of several genes in
apoptosis pathways with thymoquinone treatment in estrogen
receptor-positive breast cancer cells.
Abstract: MicroRNAs are an important class of gene expression
regulators that are involved in many biological processes including
embryogenesis. miR-125b is a conserved microRNA that is enriched
in the nervous system. We have previously reported the function of
miR-125b in neuronal differentiation of human cell lines. We also
discovered the function of miR-125b in regulating p53 in human and
zebrafish. Here we further characterize the brain defects in zebrafish
embryos injected with morpholinos against miR-125b. Our data
confirm the essential role of miR-125b in brain morphogenesis
particularly in maintaining the balance between proliferation, cell
death and differentiation. We identified lunatic fringe (lfng) as an
additional target of miR-125b in human and zebrafish and suggest
that lfng may mediate the function of miR-125b in neurogenesis.
Together, this report reveals new insights into the function of miR-
125b during neural development of zebrafish.
Abstract: Transcription factor p53 has a powerful tumor
suppressing function that is associated with many cancers. However,
p53 of the molecular weight was higher make the limitation across to
skin or cell membrane. Thymidine dinucleotide (pTT), an
oligonucleotide, can activate the p53 transcription factor. pTT is a
hydrophilic and negative charge oligonucleotide, which delivery in to
cell membrane need an appropriate carrier. The aim of this study was
to improve the bioavailability of the nucleotide fragment, thymidine
dinucleotide (pTT), using elasic liposome carriers to deliver the drug
into the skin. The study demonstrate that dioleoylphosphocholine
(DOPC) incorporated with sodium cholate at molar ratio 1:1 can
archived the particle size about 220 nm. This elastic liposome could
penetration through skin from stratum corneum to whole epidermis by
confocal laser scanning microscopy (CLSM). Moreover, we observed
the the slight increase in generation of p53 by western blot.
Abstract: The p53 tumor suppressor gene plays two important
roles in genomic stability: blocking cell proliferation after DNA
damage until it has been repaired, and starting apoptosis if the
damage is too critical. Codon 72 exon4 polymorphism (Arg72Pro) of
the P53 gene has been implicated in cancer risk. Various studies have
been done to investigate the status of p53 at codon 72 for arginine
(Arg) and proline (Pro) alleles in different populations and also the
association of this codon 72 polymorphism with various tumors. Our
objective was to investigate the possible association between P53
Arg72Pro polymorphism and susceptibility to colorectal cancer
among Isfahan and Chaharmahal Va Bakhtiari (a part of south west
of Iran) population. We investigated the status of p53 at codon 72 for
Arg/Arg, Arg/Pro and Pro/Pro allele polymorphisms in blood
samples from 145 colorectal cancer patients and 140 controls by
Nested-PCR of p53 exon 4 and digestion with BstUI restriction
enzyme and the DNA fragments were then resolved by
electrophoresis in 2% agarose gel. The Pro allele was 279 bp, while
the Arg allele was restricted into two fragments of 160 and 119 bp.
Among the 145 colorectal cancer cases 49 cases (33.79%) were
homozygous for the Arg72 allele (Arg/Arg), 18 cases (12.41%) were
homozygous for the Pro72 allele (Pro/Pro) and 78 cases (53.8%)
found in heterozygous (Arg/Pro).
In conclusion, it can be said that p53Arg/Arg genotype may be
correlated with possible increased risk of this kind of cancers in south
west of Iran.