Cooperative inhibitory effects of antisense oligonucleotide of cell adhesion molecules and cimetidine on cancer cell adhesion.
OBJECTIVE
To explore the cooperative effects of antisense oligonucleotide (ASON) of cell adhesion molecules and cimetidine on the expression of E-selectin and ICAM-1 in endothelial cells and their adhesion to tumor cells.


METHODS
After treatment of endothelial cells with ASON and/or cimetidine and induction with TNF-alpha, the protein and mRNA changes of E-selectin and ICAM-1 in endothelial cells were examined by flow cytometry and RT-PCR, respectively. The adhesion rates of endothelial cells to tumor cells were measured by cell adhesion experiment.


RESULTS
In comparison with TNF-alpha inducing group, lipo-ASON and lipo-ASON/cimetidine could significantly decrease the protein and mRNA levels of E-selectin and ICAM-1 in endothelial cells, and lipo-ASON/cimetidine had most significant inhibitory effect on E-selectin expression (from 36.37+/-1.56% to 14.23+/-1.07%, P<0.001). Meanwhile, cimetidine alone could inhibit the expression of E-selectin(36.37+/-1.56% vs 27.2+/-1.31%, P<0.001), but not ICAM-1 (69.34+/-2.50% vs 68.07+/-2.10%,P>0.05) and the two kinds of mRNA, either. Compared with TNF-alpha inducing group, the rate of adhesion was markedly decreased in lipo-E-selectin ASON and lipo-E-selectin ASON/cimetidine treated groups(P<0.05), and lipo-E-selectin ASON/cimetidine worked better than lipo-E-selectin ASON alone except for HepG2/ECV304 group (P<0.05). However, the decrease of adhesion was not significant in lipo-ICAM-1 ASON and lipo-ICAM-1 ASON/cimetidine treated groups except for HepG2/ECV304 group (P>0.05).


CONCLUSIONS
These data demonstrate that ASON in combination with cimetidine in vitro can significantly reduce the adhesion between endothelial cells and hepatic or colorectal cancer cells, which is stronger than ASON or cimetidine alone. This study provides some useful proofs for gene therapy of antiadhesion.
Tang
Nan-Hong
NH
Hepato-Biliary Surgery Institute of Fujian Province, Union Hospital, 29 Xinquan Road, Fuzhou 350001, Fujian Province, China. fztnh@sina.com
Chen
Yan-Ling
YL
Wang
Xiao-Qian
XQ
Li
Xiu-Jin
XJ
Yin
Feng-Zhi
FZ
Wang
Xiao-Zhong
XZ
D002448:Cell Adhesion / Q000187:drug effects; D045744:Cell Line, Tumor; D002927:Cimetidine / Q000494:pharmacology*; D015179:Colorectal Neoplasms; D019040:E-Selectin / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D004727:Endothelium / Q000166:cytology; D015870:Gene Expression; D006635:Histamine H2 Antagonists / Q000494:pharmacology*; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics*; D008113:Liver Neoplasms; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction
D019040:E-Selectin; D006635:Histamine H2 Antagonists; D016376:Oligonucleotides, Antisense; D018799:Intercellular Adhesion Molecule-1; D002927:Cimetidine