The RGD Domain of Human Osteopontin Promotes Tumor Growth and Metastasis through Activation of Survival Pathways |
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| Authors: | Donald Courter Hongbin Cao Shirley Kwok Christina Kong Alice Banh Peiwen Kuo Donna M. Bouley Carmen Vice Odd Terje Brustugun Nicholas C. Denko Albert C. Koong Amato Giaccia Quynh-Thu Le |
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| Affiliation: | 1. Department of Radiation Oncology, Stanford University, Stanford, California, United States of America.; 2. Department of Pathology, Stanford University, Stanford, California, United States of America.; 3. Department of Comparative Medicine, Stanford University, Stanford, California, United States of America.; 4. Department of Oncology, Oslo University Hospital – Norwegian Radium Hospital, Oslo, Norway.;Roswell Park Cancer Institute, United States of America |
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| Abstract: | BackgroundHuman osteopontin (OPN), a known tumor associated protein, exists in different isoforms, whose function is unclear. It also possesses a RGD domain, which has been implicated in diverse function. Here, we use genetic approaches to systematically investigate the function of the RGD domain in different OPN isoforms on tumor progression and metastasis for 2 different solid tumor models.Methodology/Principal FindingsUsing isoform-specific qRT-PCR, we found that OPN-A and B were the main isoforms overexpressed in evaluated human tumors, which included 4 soft tissue sarcomas, 24 lung and 30 head and neck carcinomas. Overexpression of either OPN-A or B in two different cell types promoted local tumor growth and lung metastasis in SCID mouse xenografts. However, expression of either isoform with the RGD domain either mutated or deleted decreased tumor growth and metastasis, and resulted in increased apoptosis by TUNEL staining. In vitro, whereas mutation of the RGD domain did not affect cell-cell adhesion, soft agar growth or cell migration, it increased apoptosis under hypoxia and serum starvation. This effect could be mitigated when the RGD mutant cells were treated with condition media containing WT OPN. Mechanistically, the RGD region of OPN inhibited apoptosis by inducing NF-κB activation and FAK phosphorylation. Inhibition of NF-κB (by siRNA to the p65 subunit) or FAK activation (by a inhibitor) significantly increased apoptosis under hypoxia in WT OPN cells, but not in RGD mutant cells.Conclusion/SignificanceUnlike prior reports, our data suggest that the RGD domain of both OPN-A and B promote tumor growth and metastasis mainly by protecting cells against apoptosis under stressed conditions and not via migration or invasion. Future inhibitors directed against OPN should target multiple isoforms and should inhibit cell survival mechanisms that involve the RGD domain, FAK phosphorylation and NF-κB activation. |
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