Marine-derived oligosaccharide sulfate (JG3) suppresses heparanase-driven cell adhesion events in heparanase over-expressing CHO-K1 cells.
OBJECTIVE
To elucidate the detailed mechanisms underlying the appreciable effects of JG3, a novel marine-derived oligosaccharide, on cell migration using a Chinese hamster ovary (CHO) cell line stably over-expressing heparanase.


METHODS
A retrovirus infection system was used to establish a CHO-K1 cell line stably transfected with heparanase. Immunocytochemistry was used to assess cell morphology. Flow cytometry was selected to analyze the activation of beta1-integrin, and Western blotting was used to analyze the downstream effects on the cell adhesion pathway. An affinity precipitation assay was used to determine activation of the small GTPases, Rac1, and RhoA.


RESULTS
JG3 abolished heparanase-driven formation of focal adhesions and cell spreading. Although JG3 failed to block the heparanase-triggered activation of beta1-integrin or the phosphorylation of Src, the oligosaccharide caused a significant dephosphorylation of FAK and subsequent inactivation of Erk. Furthermore, JG3 was found to arrest the activation of Rac1.


CONCLUSIONS
All these findings help form an alternative view to understand the mechanisms underlying the inhibitory effects of JG3 on cell motility.Acta Pharmacologica Sinica (2009) 30: 1033-1038; doi: 10.1038/aps.2009.97; published online 22 June 2009.
Li
Qiu-Ning
QN
Division of Anti-Tumor Pharmacology, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 201203, China.
Liu
Hai-Ying
HY
Xin
Xian-Liang
XL
Pan
Qiu-Ming
QM
Wang
Lu
L
Zhang
Jing
J
Chen
Qin
Q
Geng
Mei-Yu
MY
Ding
Jian
J
D000818:Animals; D016466:CHO Cells / Q000187:drug effects / Q000502:physiology; D002448:Cell Adhesion / Q000187:drug effects* / Q000502:physiology; D002465:Cell Movement / Q000187:drug effects* / Q000502:physiology; D006224:Cricetinae; D003412:Cricetulus; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism; D008351:Mannans / Q000737:chemistry / Q000494:pharmacology*; D015394:Molecular Structure; D009362:Neoplasm Metastasis; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D020830:rac1 GTP-Binding Protein / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism; D000099175:Heparanase
D005966:Glucuronidase; D019012:Integrin beta1; D008351:Mannans; D020830:rac1 GTP-Binding Protein; D020742:rhoA GTP-Binding Protein; D000099175:Heparanase; C524388:oligomannurarate sulfate