Acta Pharmacologica Sinica (2009) 30: 1033-1038; doi: 10.1038/aps.2009.97

Acta Pharmacologica Sinica (2009) 30: 1033-1038; doi: 10.1038/aps.2009.97; published online 22 June 2009

Original Article

Marine-derived oligosaccharide sulfate (JG3) suppresses heparanase-driven cell adhesion events in heparanase over-expressing CHO-K1 cells

Qiu-ning LI, Hai-ying LIU, Xian-liang XIN, Qiu-ming PAN, Lu WANG, Jing ZHANG, Qin CHEN, Mei-yu GENG^*, Jian DING^*

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

Aim: 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 β1-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 β1-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.

Conclusion: All these findings help form an alternative view to understand the mechanisms underlying the inhibitory effects of JG3 on cell motility.

Keywords: JG3; heparanase; cell adhesion; cell motility

This work was supported by Natural Science Foundation of China for Distinguished Young Scholars (30725046) and the Natural Science Foundation of China for Innovation Research Group (30721005).

* To whom correspondence should be addressed.
Email mygeng@mail.shcnc.ac.cn (Mei-yu GENG), suozhang@mail.shcnc.ac.cn (Jian DING)
Received 2009-05-07 Accepted 2009-05-20

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