Using siRNA to Uncover Novel Oncogenic Signaling Pathways

  • Jin-Mei Lai
  • Chi-Ying F. Huang
  • Chang-Han Chen
Part of the Methods in Molecular Biology book series (MIMB, volume 623)


Tumor invasion and metastasis are the primary causes of cancer patient mortality, underscoring the need for identification of novel genes and signaling pathways that mediate these prognosis-determining phenomena. To identify and characterize novel lung adenocarcinoma genes associated with lung cancer progression, we created a bioinformatics-based approach that focuses on human cell-cycle-regulated genes that have evolved only in higher organisms but not in lower eukaryotic cells. In siRNA experiments in lung cancer cells, FLJ10540 was identified as one of several novel targets involved in cell migration and invasion. Here, we demonstrate that PI3K inhibition affects FLJ10540-mediated cell migration and invasion and further, that FLJ10540 knockdown ablates AKT-Ser473 phosphorylation. Taken together, these findings indicate that the FLJ10540/PI3K/AKT pathway may harbor new therapeutic targets for treating invasive lung adenocarcinoma.

Key words

FLJ10540 Migration Invasion PI3K/ AKT VEGF-A 


  1. 1.
    Friedl, P., and Wolf, K. (2003) Tumour-cell invasion and migration: diversity and escape mechanisms. Nat. Rev. Cancer 3, 362–374.CrossRefPubMedGoogle Scholar
  2. 2.
    Osaki, M., Oshimura, M., and Ito, H. (2004) PI3K-Akt pathway: its functions and alterations in human cancer. Apoptosis 9, 667–676.CrossRefPubMedGoogle Scholar
  3. 3.
    Ramesh, R., Ito, I., Gopalan, B., Saito, Y., Mhashilkar, A. M., and Chada, S. (2004) Ectopic production of MDA-7/IL-24 inhibits invasion and migration of human lung cancer cells. Mol. Ther. 9, 510–518.CrossRefPubMedGoogle Scholar
  4. 4.
    Su, L. J., Chang, C. W., Wu, Y. C., Chen, K. C., Lin, C. J., Liang, S. C., et al. (2007) Selection of DDX5 as a novel internal control for Q-RT-PCR from microarray data using a block bootstrap re-sampling scheme. BMC Genomics 8, 140.CrossRefPubMedGoogle Scholar
  5. 5.
    Chen, C. H., Lai, J. M., Chou, T. Y., Chen, C. Y., Su, L. J., Lee, Y. C., et al. (2009) VEGFA upregulates FLJ10540 and modulates migration and invasion of lung cancer via PI3K/AKT pathway. PLoS ONE 4, e5052.CrossRefPubMedGoogle Scholar
  6. 6.
    Chen, C. H., Lu, P. J., Chen, Y. C., Fu, S. L., Wu, K. J., Tsou, A. P., et al. (2007) FLJ10540-elicited cell transformation is through the activation of PI3-kinase/AKT pathway. Oncogene 26, 4272–4283.CrossRefPubMedGoogle Scholar
  7. 7.
    Cantley, L. C. (2002) The phosphoinositide 3-kinase pathway. Science 296, 1655–1657.CrossRefPubMedGoogle Scholar
  8. 8.
    Zachary, I. (2003) VEGF signalling: integration and multi-tasking in endothelial cell biology. Biochem. Soc. Trans. 31, 1171–1177.CrossRefPubMedGoogle Scholar
  9. 9.
    Laramée, M., Chabot, C., Cloutier, M., Stenne, R., Holgado-Madruga, M., Wong, A. J., and Royal, I. (2007) The scaffolding adapter Gab1 mediates vascular endothelial growth factor signaling and is required for endothelial cell migration and capillary formation. J. Biol. Chem. 282, 7758–7769.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Jin-Mei Lai
    • 1
  • Chi-Ying F. Huang
    • 2
  • Chang-Han Chen
    • 3
    • 4
  1. 1.Department of Life ScienceFu-Jen Catholic UniversityTaipeiTaiwan
  2. 2.Institute of Clinical MedicineNational Yang-Ming UniversityTaipeiTaiwan
  3. 3.Department of OtolaryngologyChang Gung MemorialHospital-Kaohsiung Medical Center, Chang Gung University College of MedicineKaohsiungTaiwan
  4. 4.Kaohsiung Chang Gung Head and Neck Oncology GroupChang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University College of MedicineKaohsiungTaiwan

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