Activation of STAT3 signaling in human stomach adenocarcinoma drug-resistant cell line and its relationship with expression of vascular endothelial growth factor.
OBJECTIVE
To investigate the difference in activation of STAT3 signaling between two human stomach adenocarcinoma cell lines: 5-fluorouracil resistant cell line and its parental cell line, and to evaluate its relationship with the expression of vascular endothelial growth factor (VEGF).


METHODS
Western blot and electrophoretic mobility shift assay (EMSA) were used to detect the expression of phospho-STAT3 protein and constitutive activation of STAT3 in two human stomach adenocarcinoma cell lines, 5-fluorouracil resistant cell line SGC7901/R and its parental cell line SGC7901, respectively. The mRNA expression of VEGF was analysed by semi-quantitative RT-PCR. The expressive intensity of VEGF protein was measured by immunocytochemistry.


RESULTS
The expressions of phospho-STAT3 protein and constitutive activation of STAT3 between two human stomach adenocarcinoma cell lines were different. Compared with the parental cell line SGC7901, the STAT3-DNA binding activity and the expressive intensity of phospho-STAT3 protein were lower in the drug-resistant cell line SGC7901/R. The expression levels of VEGF mRNA and its encoded protein were also decreased in drug-resistant cell line.


CONCLUSIONS
Over-expression of VEGF may be correlated with elevated STAT3 activation in parental cell line. Lower VEGF expression may be correlated with decreased STAT3 activation in resistant cell line, which may have resulted from negative feedback regulation of STAT signaling.
Yu
Li-Fen
LF
Department of Gastroenterology, Ruijin Hospital of Shanghai Second Medical University, 197 Ruijin er Road, Shanghai 200025, China. graceyu1028@sohu.com
Cheng
Ying
Y
Qiao
Min-Min
MM
Zhang
Yong-Ping
YP
Wu
Yun-Lin
YL
D000230:Adenocarcinoma; D015153:Blotting, Western; D045744:Cell Line, Tumor / Q000187:drug effects / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D019008:Drug Resistance, Neoplasm / Q000502:physiology; D025461:Feedback, Physiological; D006801:Humans; D010766:Phosphorylation; D012333:RNA, Messenger / Q000032:analysis; D050796:STAT3 Transcription Factor; D015398:Signal Transduction / Q000502:physiology; D013274:Stomach Neoplasms; D015534:Trans-Activators / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*
D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D015534:Trans-Activators; D042461:Vascular Endothelial Growth Factor A; D004247:DNA