GATA-3 augmentation down-regulates Connexin43 in Helicobacter pylori associated gastric carcinogenesis

Figures & data

Table 1. Transcriptional Factors DNA array in H. pylori associated gastric carcinogenesis (standardized value)

	H. pylori (-)	H. pylori (+)
Gene Name	NGM	NAG	CAG	IM	DYS	GC
AP-1	0.364475	0.236852	0.437483	0.109007	0.33254	0.557029
CdxA/NKX2	0.025694	0.162097	0.303745	0.077073	0.23202	0.373394
MyoD	0.755692	0.708577	0.575592	0.815667	0.69901	0.896107
c-Myc-responsive region	0.021542	0.151038	0.17115	0.155812	0.45871	0.531179
MZF1	0.130042	0.212263	0.390068	0.063666	0.70764	0.640325
myc-CF1	0.042735	0.120381	0.08333	0.065778	0.08691	0.220635
USF-1	0.59681	0.586308	0.504982	0.274977	0.68181	0.724868
Nkx-2.5	0.07258	0.189742	0.212145	0.06976	0.26695	0.160128
SRY	0.123837	0.22467	0.054424	0.18344	0.02147	0.006878
NF-1(2)	0.193751	0.20134	0.0334	0.390525	0.23373	0.499282
ADR1	0.839109	0.792412	0.768519	0.863365	0.74375	0.850454
p300	0.812017	0.802346	0.77387	0.901597	0.65291	0.594067
E12/E47	0.177671	0.206015	0.147324	0.25466	0.05314	0.457672
E2F-1	0.973607	0.909062	0.913756	1.02194	0.98074	1.004649
NF-E1 (YY1)	0.828665	0.737391	0.773008	0.838459	0.75935	0.906803
GATA-3	0.181778	0.28055	0.48277	0.59822	0.4974	0.64002
NF-Y	0.10094	0.151803	0.026937	0.045057	0.02396	0.073318
HLF	0.11418	0.1398	0.04269	0.090441	0.04606	0.065457
EVI-1	0.725453	0.385328	0.855196	0.916467	0.88423	0.898413
LyF-1	0.744593	0.750652	0.73725	0.894202	0.87372	0.895843
1-Oct	0.13087	0.12645	0.172252	0.116971	0.0952	0.181557
c-Ets-1	0.116976	0.09053	0.079786	0.307845	0.26178	0.323432
HFH-2	0.951846	0.972714	1.017406	1.064722	1.05287	1.053023

Figure 1. Comparison of GATA-3 and Cx43 expression among developmental stages of H. pylori associated gastric carcinoma in clinical specimens. (A, B) Quantitative RT-PCR analysis of GATA-3 (A) and Cx43 (B) in H. pylori (-) NGM, H. pylori (+) CAG and GC patients (n=15). *** p < 0.001. (C) Representative images of GATA-3 and Cx43 protein levels in H. pylori (-) NGM, H. pylori (-/+) CAG, IM and GC.

Figure 2. The effects of H. pylori on GATA-3 and Cx43 expressions in GES-1 cells. (A, B) Quantitative RT-PCR analysis of GATA-3 (A) and Cx43 (B) in GES-1 cells with H. pylori 24 and 48 hours infections. ** p < 0.01; *** p < 0.001. (C) Morphology of normal GES-1 cells and 24/48 hours H. pylori infected GES-1 cells. (D) Western blot analysis of GATA-3 and Cx43 protein levels in GES-1 cells under 24 and 48 hours H. pylori infections.

Figure 3. GATA-3 negatively regulates Cx43 by directly binding to its promoters. (A, B) Quantitative RT-PCR analysis of GATA-3 (A) and Cx43 (B) levels in H. pylori infected GES-1 cells treated with a scramble control, GATA-3 siRNA-1, GATA-3 siRNA-2. (C) Western blot analysis of GATA-3 and Cx43 protein levels in H. pylori infected GES-1 cells treated with a scramble control, GATA-3 siRNA-1, and GATA-3 siRNA-2. (D) GATA-3 direct binds to Cx43 promoters. HEK-293T cells co-transfected with GATA-3 and Cx43-pGL3 plasmids. The results are displayed as the ratio of firefly luciferase activity in the pGL3-Cx43-GATA-3-transfected cells to the activity in the pGL3-Cx43 controls. *** p < 0.001.

Figure 4. The effects of H. pylori on GATA-3 and Cx43 expression in Mongolian gerbils. (A) The alterative GATA-3 and Cx43 mRNA in Mongolian gerbils with H. pylori 48-week infection (n=10) compared to control group (n=10). *** p < 0.001. (B) The correlation of GATA-3 and Cx43 mRNA levels from Mongolian gerbils (10 from experimental group and 10 from control group). (C) Profile of Mongolian gerbil (upper left); typical macroscopic manifestation of normal stomach in control group (top center) and inflammatory edema in experimental group (upper right); histologic sections of normal gastric mucosa (left bottom), H. pylori induced severe non-atrophic gastritis (bottom center), and H. pylori induced atrophic gastritis (right bottom).