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Cancer qPCR array + QuantiMir Poster Citation:
#133 Aryl
hydrocarbon receptor agonists inhibit breast cancer cell growth
and modulate microRNA expression. Shu
Zhang1,
Craig Rowlands2, Stephen Safe1. 1Texas
A&M University, College Station, TX; 2The Dow
Chemical Company, Midland, MI.
MDA-MB-468 and BT474 are estrogen receptor (ER)-negative
and ER-positive breast cancer cell lines, respectively, that
also express the aryl hydrocarbon receptor (AhR). Treatment of
these cell lines with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD),
1,2,3,7,8-pentachlorodibenzo-p-dioxin (PCDD),
2,3,7,8-tetrachlorodibenzofuran (TCDF),
2,3,4,7,8-pentachlorodibenzofuran (PCDF),
3,3’,4,4’,5-pentachlorobiphenyl (PCB), and the selective AhR
modulator (SAhRM) 6-methyl-1,3,8-trichlorodibenzofuran (MCDF)
induced CYP1A1 expression, a typical AhR-responsive gene. In
addition, treatment with 40 nM TCDD, PCDD, TCDF and PCDF, 100 nM
PCB and 5 uM MCDF resulted in a >30-80% and 25-50% decreased in
MDA-MB-468 and BT474 cell proliferation, respectively. Moreover,
in cells treated with the halogenated aromatic TCDD, PCDD, TCDF,
PCDF and PCB congeners, the growth inhibitory effects were
completely abrogated after cotransfection with a small
inhibitory RNA for the AhR (iAhR). In contrast, iAhR only
partially reversed the antiproliferative effects of 6-MCDF,
suggesting that this compound activates both AhR-dependent and
AhR-independent growth inhibitory pathways.
The potential role of
microRNAs in mediating the effects of these compounds was
investigated using a System
Biosciences QuantiMir Cancer MicroRNA qPCR Array. The
results show that 6-MCDF and the five halogenated aromatics
modulate expression of several common microRNAs including
let-7-family, miR-103, miR-198 and miR-373. However, it was also
evident that 6-MCDF and the five halogenated aromatics
differentially modulated microRNA expression, and current
studies are focused on identifying and characterizing
structure-dependent activation of microRNAs via AhR-dependent
and independent pathways. [Supported by the Dow Chemical Company
and National Institutes of Health (ES09106 and ES04917)]
QuantiMir Poster Citation:
#5064 Identification of differentially expressed miRNAs in
gastric cancer. Oleg
Tchernitsa1, Reinhold Schafer1, Balazzs
Gyorffy1, Ulf Neumann1, Matthias P.A.
Ebert2, Christoph Rocken1. 1Charite
University Hospital, Berlin, Germany; 2Technical
University Munich, Munich, Germany.
Aims: In this study we investigated the differential
expression of microRNA (miRNA) in lesional and nonlesional
samples of gastric cancer patients.
Materials and Methods: Samples from gastric cancer and
corresponding nonneoplastic gastric mucosa were obtained from
six patients with intestinal type gastric cancer (m:f=4:1).
Tissue samples were obtained immediately after surgery and
stored at -80 °C. MiRNA was purified using the mirVana™ qRT-PCR
miRNA Detection Kit (Ambion). Oligonucletide miRNA specific
microarrays were spotted in the Laboratory for Functional
Genetics of Charité (O.T., R.S.) using Invitrogen NCode™
Multi-Species miRNA Microarray Probe Set containing 857
mammalian probes. All probes were spotted with two replica on
epoxy treated slides from Corning. Labeling was done using
Invitrogen NCode™ miRNA labeling system. Microarray images were
obtained on an Agilent G2565AA scanner at 10 μm resolution.
Image analysis was performed using ImaGene software (BioDiscovery,
Inc., El Segundo, USA). miRNA profiles of paired normal and
tumor samples were compared. Statistical evaluation was
performed using the BRB-ArrayTools version 3.6.0 beta 1.
Differential expression was validated by RT-PCR using
QuantiMir System (SBI System Biosciences).
Results: Thirty-three miRNAs were identified by Significance
Analysis of Microarrays (SAM) analysis as up-regulated in
gastric carcinoma compared to adjacent normal tissue. Fourteen
miRNAs were also confirmed by Class Comparison analysis. All
fourteen up-regulated miRNA transcripts were subsequently
validated by QuantiMir RT-PCR
and included hsa_miR_30b, hsa_let_7a, hsa_miR_7, hsa_miR_145,
hsa_let_7b, hsa_let_7f, hsa_miR_93, hsa_let_7c, hsa_miR_125b,
hsa_miR_320, hsa_miR_18a, hsa_miR_214, hsa_miR_23b,
hsa_miR_106b. SAM analysis did not identify any commonly
down-regulated miRNA in tumor tissues.
Conclusion: Intestinal type gastric cancer can be added to the
growing list of tumors with differentially expressed miRNAs.
Future studies will have to show whether differential expression
of miRNA in gastric cancer is involved in tumor development and
progression and is hence a putative target for diagnostic,
prognostic or therapeutic purposes
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