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W. Xie
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P1.04 - Poster Session/ Biology, Pathology, and Molecular Testing (ID 233)
- Event: WCLC 2015
- Type: Poster
- Track: Biology, Pathology, and Molecular Testing
- Presentations: 1
- Moderators:
- Coordinates: 9/07/2015, 09:30 - 17:00, Exhibit Hall (Hall B+C)
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P1.04-077 - KIF5B-RET Fusion Kinase Promotes Cell Invasion and Migration Which Can Be Suppressed by RET Inhibitors (ID 2512)
09:30 - 09:30 | Author(s): W. Xie
- Abstract
Background:
Non-small cell lung cancer has become the leading cause of cancer-related deaths worldwide. The subset of NSCLC can be further defined at molecular level by driver mutations that occur in multiple oncogenes, such as EGFR, KRAS and EML4/ALK alterations. The KIF5B-RET fusion gene has been established as a new oncogenic driver in NSCLC. Several studies have demonstrated that KIF5B-RET promote cell growth and tumorigenicity, however, few progress has been made further. Our study aims to investigate other characters of KIF5B-RET fusion gene and tries to explore the potential signaling pathways involved in the gene functions.
Methods:
Lentivirus（encoding KIF5B-RET）was used to transfect the lung epithelial cell line BEAS-2B and lung cancer cell line A549 to generate stable transfectant and the protein expression was analysed using western blot . To verify the oncogenic features of KIF5B-RET in vitro, we detected its expression genetically followed by CCK8 assay, colony formation assay, transwell and Annexin V-FITC/PI double staining to explore proliferation, invasion, migration and apoptosis. The mechanism by which KIF5B-RET kinase induced invasion and migration was investigated by western blot, and administration of RET and SRC inhibitions.
Results:
The stable transfected cell line expressed phosphorylation RET, examined by western blot, suggesting that KIF5B-RET could automatically activate RET protein in the absence of ligand. Firstly we detected the basic characters of KIF5B-RET, but found no significant difference in proliferation as it’s reported in previous studies. To further detect the function of KIF5B-RET fusion gene, we focused on characters of invasion, migration, and apoptosis. We demonstrated that KIF5B-RET showed a significantly increased ability of invasion and migration compared with control group, suggesting that KIF5B-RET fusion gene could promote cell invasion and migration. However, no change was observed after treating the transfected cells with Cisplatin, indicating the gene may have no influence on apoptosis. As we all know, RET tyrosine kinase can activate ERK which belongs to the downstream signaling system. Our restult showed that KIF5B-RET fusion kinase can also induced activation of ERK and even SRC kinase. Finally, we found that stable cells became sensitive to the RET tyrosine kinase inhibitors Sunitinib and Apatinib. The invasion and migration could be suppressed by RET or SRC inhibitors significantly.
Conclusion:
Out data showed that KIF5B-RET fusion gene can activate ERK and SRC kinase through activating RET tyrosine kinase, and promote migration and invasion in vitro ，but did not have an effort on proliferation and apoptosis. RET inhibitor Apatinib and Sunitinib and SRC inhibitor could suppress the phenomenon of invasion and migration, suggesting that KIF5B-RET promotes invasion and migration through activation of SRC kinase. Our preclinical data demonstrated the antitumor activities of Apatinib and Sunitinib against KIF5B-RET gene fusion-driven cells and indicated the therapeutic potential of tyrosine kinase inhibitors targeting RET, which may benefit this certain subpopulation of NSCLC patients.
