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PI3K Blockade by Ad-PTEN Inhibits Invasion and Induces Apoptosis in Radial Growth Phase and Metastatic Melanoma Cells

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Abstract

Background

Melanoma is an aggressive tumor with a propensity to rapidly metastasize. The PTEN gene encodes a phosphatase with an unusual dual specificity for proteins and lipids. Mutations of PTEN have been found in various human cancers, including glioblastoma, prostate, breast, lung, and melanoma. Here we investigate in vitro the effects of blocking PI3K signaling using adenoviral-delivered PTEN (Ad-PTEN) in cell lines derived from both early- and late-stage melanoma.

Materials and Methods

Ad-PTEN transduced melanoma cell lines or normal cells were assayed for cell death, apoptosis, gene expression, invasion and migration, and regulation of angiogenesis.

Results

The PTEN locus from RGP and metastatic melanoma cell lines was sequenced; no coding region mutations were found. Adenoviral transfer of PTEN into melanoma cells containing wild-type PTEN alleles led to tumor-specific apoptosis and growth inhibition, with coordinate inhibition of AKT phosphorylation. Ad-PTEN suppressed cell migration by metastatic melanoma cells with concomitant increase in the level of cell surface E-cadherin. Immunohistochemical and confocal analyses localized PTEN to the cytoplasm and demonstrated enrichment at the cell membrane. Ad-PTEN inhibited angiogenesis as demonstrated by the tube formation assay using human vascular endothelial cells.

Conclusions

These studies indicate that Ad-PTEN can inhibit tumor cells via multiple mechanisms and has proapoptotic, anti-metastatic, and anti-angiogenic properties. Thus, PI3K blockade via Ad-PTEN may be a promising approach for the treatment of early- and late-stage melanoma, even in tumors that do not harbor PTEN mutations.

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References

  1. Meier F, Satyamoorthy K, Nesbit M, et al. (1998) Molecular events in melanoma development and progression. Front. Biosci. 3: 1005–1010.

    Article  Google Scholar 

  2. Saida T. (1993) Recent advances in melanoma research. J. Dermatol. Sci. 26: 1–13.

    Article  Google Scholar 

  3. Herlyn M. (1993) Molecular and Cellular Biology for Melanoma. Austin, TX: R.G. Landes Co.; pp. 1–102.

  4. Healy E, Belgaid CE, Takata M, et al. (1996) Allelotypes of primary cutaneous melanoma and benign melanocytic nevi. Cancer Res. 56: 589–593.

    PubMed  CAS  Google Scholar 

  5. Bastian BC, LeBoit PE, Hamm H, Brocker EB, Pinkel D. (1998) Chromosomal gains and losses in primary cutaneous melanomas detected by comparative genomic hybridization. Cancer Res. 58: 2170–2175.

    PubMed  CAS  Google Scholar 

  6. Thompson FH, Emerson J, Olson S, et al. (1995) Cytogenetics of 158 patients with regional or disseminated melanoma. Subset analysis of near-diploid and simple karyotypes. Cancer Genet. Cytogenet. 83: 93–104.

    Article  CAS  PubMed  Google Scholar 

  7. Herbst RA, Weiss J, Ehnis A, Cavenee WK, Arden KC. (1994) Loss of heterozygosity for 10q22-10qter in malignant melanoma progression. Cancer Res. 54: 3111–3114.

    PubMed  CAS  Google Scholar 

  8. Li J, Yen C, Liaw D, et al. (1997) PTEN, a putative protein tyrosine phosphatase gene mutated in human brain, breast, and prostate cancer. Science 275: 1943–1947.

    Article  CAS  PubMed  Google Scholar 

  9. Steck PA, Pershouse MA, Jasser SA, et al. (1997) Identification of a candidate tumour suppressor gene, MMAC1, at chromosome 10q23.3 that is mutated in multiple advanced cancers. Nat. Genet. 15: 356–362.

    Article  CAS  PubMed  Google Scholar 

  10. Li DM, Sun H. (1997) TEP1, encoded by a candidate tumor suppressor locus, is a novel protein tyrosine phosphatase regulated by transforming growth factor beta. Cancer Res. 57: 2124–2129.

    PubMed  CAS  Google Scholar 

  11. Tamura M, Gu J, Matsumoto K, Aota S, Parsons R, Yamada KM. (1998) Inhibition of cell migration, spreading, and focal adhesions by tumor suppressor PTEN. Science 280: 1614–1617.

    Article  CAS  PubMed  Google Scholar 

  12. Tamura M, Gu J, Tran H, Yamada KM. (1999) PTEN gene and integrin signaling in cancer. J. Natl. Cancer Inst. 91: 1820–1828.

    Article  CAS  PubMed  Google Scholar 

  13. Tamura M, Gu J, Danen EH, Takino T, Miyamoto S, Yamada KM. (1999) PTEN interactions with focal adhesion kinase and suppression of the extracellular matrix-dependent phosphatidylinositol 3-kinase/Akt cell survival pathway. J. Biol. Chem. 274: 20693–20703.

    Article  CAS  PubMed  Google Scholar 

  14. Tamura M, Gu J, Takino T, Yamada KM. (1999) Tumor suppressor PTEN inhibition of cell invasion, migration, and growth: differential involvement of focal adhesion kinase and p130Cas. Cancer Res. 59: 442–449.

    PubMed  CAS  Google Scholar 

  15. Guldberg P, thor Straten P, Birck A, Ahrenkiel V, Kirkin AF, Zeuthen J. (1997) Disruption of the MMAC1/PTEN gene by deletion or mutation is a frequent event in malignant melanoma. Cancer Res. 57: 3660–3663.

    PubMed  CAS  Google Scholar 

  16. Robertson GP, Furnari FB, Miele ME, et al. (1998) In vitro loss of heterozygosity targets the PTEN/MMAC1 gene in melanoma. Proc. Natl. Acad. Sci. U.S.A. 95: 9418–9423.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Tsou HC, Teng DH, Ping XL, et al. (1997) The role of MMAC1 mutations in early-onset breast cancer: causative in association with Cowden syndrome and excluded in BRCA1-negative cases. Am. J. Hum. Genet. 61: 1036–1043.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Olschwang S, Serova-Sinilnikova OM, Lenoir GM, Thomas G. (1998) PTEN germ-line mutations in juvenile polyposis coli. Nat. Genet. 18: 12–14.

    Article  CAS  PubMed  Google Scholar 

  19. Liaw D, Marsh DJ, Li J, et al. (1997) Germline mutations of the PTEN gene in Cowden disease, an inherited breast and thyroid cancer syndrome. Nat. Genet. 16: 64–67.

    Article  CAS  PubMed  Google Scholar 

  20. Cantley LC, Neel BG. (1999) New insights into tumor suppression: PTEN suppresses tumor formation by restraining the phosphoinositide 3-kinase/AKT pathway. Proc. Natl. Acad. Sci. U.S.A. 96: 4240–4245.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Marsh DJ, Dahia PL, Zheng Z, et al. (1997) Germline mutations in PTEN are present in Bannayan-Zonana syndrome. Nat. Genet. 16: 333–334.

    Article  CAS  PubMed  Google Scholar 

  22. Podsypanina K, Ellenson LH, Nemes A, et al. (1999) Mutation of Pten/Mmac1 in mice causes neoplasia in multiple organ systems. Proc. Natl. Acad. Sci. U.S.A. 96: 1563–1568.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Suzuki A, de la Pompa JL, Stambolic V, et al. (1998) High cancer susceptibility and embryonic lethality associated with mutation of the PTEN tumor suppressor gene in mice. Curr. Biol. 8: 1169–1178.

    Article  CAS  PubMed  Google Scholar 

  24. Christofori G, Semb H. (1999) The role of the cell-adhesion molecule E-cadherin as a tumour-suppressor gene. Trends Biochem. Sci. 24: 73–76.

    Article  CAS  PubMed  Google Scholar 

  25. Rasheed BK, Stenzel TT, McLendon RE, et al. (1997) PTEN gene mutations are seen in high-grade but not in low-grade gliomas. Cancer Res. 57: 4187–4190.

    PubMed  CAS  Google Scholar 

  26. Marsh DJ, Coulon V, Lunetta KL, et al. Mutation spectrum and genotype-phenotype analyses in Cowden disease and Bannayan-Zonana syndrome, two hamartoma syndromes with germline PTEN mutation. Hum. Mol. Genet. 7: 507–515.

  27. Maehama T, Dixon JE. (1998) The tumor suppressor, PTEN/MMAC1, dephosphorylates the lipid second messenger, phosphatidylinositol 3,4,5-trisphosphate. J. Biol. Chem. 273: 13375–13378.

    Article  CAS  PubMed  Google Scholar 

  28. Myers MP, Pass I, Batty IH, et al. (1998) The lipid phosphatase activity of PTEN is critical for its tumor suppressor function. Proc. Natl. Acad. Sci. U.S.A. 95: 13513–13518.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Sun H, Lesche R, Li DM, et al. (1999) PTEN modulates cell cycle progression and cell survival by regulating phosphatidylinositol 3,4,5,-trisphosphate and Akt/protein kinase B signaling pathway. Proc. Natl. Acad. Sci. U.S.A. 96: 6199–6204.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Stambolic V, Suzuki A, de la Pompa JL, et al. (1998) Negative regulation of PKB/Akt-dependent cell survival by the tumor suppressor PTEN. Cell 95: 29–39.

    Article  CAS  PubMed  Google Scholar 

  31. Ramaswamy S, Nakamura N, Vazquez F, et al. Regulation of G1 progression by the PTEN tumor suppressor protein is linked to inhibition of the phosphatidylinositol 3-kinase/Akt pathway. Proc. Natl. Acad. Sci. U.S.A. 96: 2110–2115.

  32. Li J, Simpson L, Takahashi M, et al. (1998) The PTEN/MMAC1 tumor suppressor induces cell death that is rescued by the AKT/protein kinase B oncogene. Cancer Res. 58: 5667–5672.

    PubMed  CAS  Google Scholar 

  33. Li DM, Sun H. (1998) PTEN/MMAC1/TEP1 suppresses the tumorigenicity and induces G1 cell cycle arrest in human glioblastoma cells. Proc. Natl. Acad. Sci. U.S.A. 95: 15406–15411.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Francis GM, Krohn EG, Woods KV, Buzaid AC, Grimm EA. (1996) Interleukin-6 production and secretion in human melanoma cell lines: regulation by interleukin-1. Melanoma Res. 6: 191–201.

    Article  CAS  PubMed  Google Scholar 

  35. Hanahan D, Weinberg RA. (2002) The hallmarks of cancer. Cell 100: 57–70.

    Article  Google Scholar 

  36. Huang J, Kontos CD. (2002) PTEN Modulates Vascular Endothelial Growth Factor-Mediated Signaling and Angiogenic Effects. J. Biol. Chem. 277: 10760–10766.

    Article  CAS  PubMed  Google Scholar 

  37. Furnari FB, Lin H, Huang HS, Cavenee WK. (1997) Growth suppression of glioma cells by PTEN requires a functional phosphatase catalytic domain. Proc. Natl. Acad. Sci. U.S.A. 94: 12479–12484.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Davies MA, Lu Y, Sano T, et al. (1998) Adenoviral transgene expression of MMAC/PTEN in human glioma cells inhibits Akt activation and induces anoikis. Cancer Res. 58: 5285–5290.

    PubMed  CAS  Google Scholar 

  39. Davies MA, Koul D, Dhesi H, et al. (1999) Regulation of Akt/PKB activity, cellular growth, and apoptosis in prostate carcinoma cells by MMAC/PTEN. Cancer Res. 59: 2551–2556.

    PubMed  CAS  Google Scholar 

  40. Davies MP, Gibbs FE, Halliwell N, et al. (1999) Mutation in the PTEN/MMAC1 gene in archival low grade and high grade gliomas. Br. J. Cancer 79: 1542–1548.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Cheney IW, Johnson DE, Vaillancourt MT, et al. (1998) Suppression of tumorigenicity of glioblastoma cells by adenovirus-mediated MMAC1/PTEN gene transfer. Cancer Res. 58: 2331–2334.

    PubMed  CAS  Google Scholar 

  42. Minaguchi T, Mori T, Kanamori Y, et al. (1999) Growth suppression of human ovarian cancer cells by adenovirus-mediated transfer of the PTEN gene. Cancer Res. 59: 6063–6067.

    PubMed  CAS  Google Scholar 

  43. Sakurada A, Hamada H, Fukushige S, et al. (1999) Adenovirus-mediated delivery of the PTEN gene inhibits cell growth by induction of apoptosis in endometrial cancer. Int. J. Oncol. 15: 1069–1074.

    PubMed  CAS  Google Scholar 

  44. Zhu X, Kwon CH, Schlosshauer PW, Ellenson LH, Baker SJ. (2001) PTEN induces G(1) cell cycle arrest and decreases cyclin D3 levels in endometrial carcinoma cells. Cancer Res. 61: 4569–4575.

    PubMed  CAS  Google Scholar 

  45. Lu Y, Lin YZ, LaPushin R, et al. (1999) The PTEN/MMAC1/TEP tumor suppressor gene decreases cell growth and induces apoptosis and anoikis in breast cancer cells. Oncogene 18: 7034–7045.

    Article  CAS  PubMed  Google Scholar 

  46. Weng LP, Smith WM, Dahia PL, et al. (1999) PTEN suppresses breast cancer cell growth by phosphatase activity-dependent G1 arrest followed by cell death. Cancer Res. 59: 5808–5814.

    PubMed  CAS  Google Scholar 

  47. Weng LP, Gimm O, Kum JB, et al. (2001) Transient ectopic expression of PTEN in thyroid cancer cell lines induces cell cycle arrest and cell type-dependent cell death. Hum. Mol. Genet. 10: 251–258.

    Article  CAS  PubMed  Google Scholar 

  48. Tanaka M, Koul D, Davies MA, Liebert M, Steck PA, Grossman HB. (2000) MMAC1/PTEN inhibits cell growth and induces chemosensitivity to doxorubicin in human bladder cancer cells. Oncogene 19: 5406–5412.

    Article  CAS  PubMed  Google Scholar 

  49. Gimm O, Perren A, Weng LP, et al. (2000) Differential nuclear and cytoplasmic expression of PTEN in normal thyroid tissue, and benign and malignant epithelial thyroid tumors. Am. J. Pathol. 156: 1693–1700.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Perren A, Komminoth P, Saremaslani P, et al. (2000) Mutation and expression analyses reveal differential subcellular compartmentalization of PTEN in endocrine pancreatic tumors compared to normal islet cells. Am. J. Pathol. 157: 1097–1103.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Ding Y, Shimada Y, Kano M, et al. (2000) PTEN/MMAC1 expression in esophageal squamous cell carcinomas. Int. J. Oncol. 17: 695–699.

    PubMed  CAS  Google Scholar 

  52. Sano T, Lin H, Chen X, et al. (1999) Differential expression of MMAC/PTEN in glioblastoma multiforme: relationship to localization and prognosis. Cancer Res. 59: 1820–1824.

    PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by Introgen Therapeutics Inc., Houston, Texas, and NCI grant CA86587 to S. C. and NCI grant CA 89778 to E. A. G.

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  1. Authors contributed equally.

Authors and Affiliations

  1. Department of Research, Introgen Therapeutics, Inc., 2250 Holcombe Blvd., Houston, Texas, 77030, USA

    Alexis L. Stewart, Abner M. Mhashilkar, Xiaohong Helena Yang, Kerry Sieger, Robert Schrock, Eric Onishi, Xin Swanson, Lou Zumstein & Sunil Chada

  2. Department of Bioimmunotherapy, The University of Texas M. D. Anderson Cancer Center, Houston, Texas, USA

    Suhendan Ekmekcioglu, John B. Mumm, Elizabeth A. Grimm & Sunil Chada

  3. Department of Thoracic and Cardiovascular Surgery, The University of Texas M. D. Anderson Cancer Center, Houston, Texas, USA

    Yuji Saito, Jack A. Roth & Rajagopal Ramesh

  4. Imperial Cancer Research Fund, Applied Development Laboratory, London, UK

    Graham J. Watson & David Snary

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Correspondence to Sunil Chada.

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Stewart, A.L., Mhashilkar, A.M., Yang, X.H. et al. PI3K Blockade by Ad-PTEN Inhibits Invasion and Induces Apoptosis in Radial Growth Phase and Metastatic Melanoma Cells. Mol Med 8, 451–461 (2002). https://doi.org/10.1007/BF03402025

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