DOI QR코드

DOI QR Code

Knockdown of Radixin by RNA interference Suppresses the Growth of Human Pancreatic Cancer Cells in Vitro and in Vivo

  • Chen, Shu-Dong (Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University) ;
  • Song, Mao-Min (Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University) ;
  • Zhong, Zhi-Qiang (Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University) ;
  • Li, Na (Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University) ;
  • Wang, Pi-Lin (Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University) ;
  • Cheng, Shi (Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University) ;
  • Bai, Ri-Xing (Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University) ;
  • Yuan, Hui-Sheng (Department of General Surgery, Beijing Tiantan Hospital, Capital Medical University)
  • Published : 2012.03.31

Abstract

Radixin, encoded by a gene on chromosome 11, plays important roles in cell motility, invasion and tumor progression. However, its function in pancreatic cancer remains elusive. In this study, radixin gene expression was suppressed with a lentivirus-mediated short-hairpin RNA (shRNA) method. We found that radixin shRNA caused down-regulation of radixin in PANC-1 cells, associated with inhibition of pancreatic cancer cell proliferation, survival, adhesion and invasive potential in vitro. When radixin-silenced cells were implanted in nude mice, tumor growth and microvessel density were significantly inhibited as compared to blank control cells or nonsense shRNA control cells. Thrombospondin-1 (TSP-1) and E-cadherin were up-regulated in radixin-silenced PANC-1 cells. Our results suggest that radixin might play a critical role in pancreatic cancer progression, possibly through invvolvement of down-regulation of TSP-1 and E-cadherin expression.

References

  1. Albini A, Iwamoto Y, Kleinman HK, et al (1978). A rapid in vitro assay for quantitating the invasive potential of tumor cells. Cancer Res, 47, 3239-45.
  2. Berx G, Staes K, van Henqel J, et al (1995). Cloning and characterization of the human invasion suppressor gene E-cadherin (CDH1). Genomics, 26, 281-9. https://doi.org/10.1016/0888-7543(95)80212-5
  3. Bretscher A, Edwards K, Fehon RG (2002). ERM proteins and merlin: integrators at the cell cortex. Nat Rev Mol Cell Biol, 3, 586-99. https://doi.org/10.1038/nrm882
  4. Butler WB, Berlinski PJ, Hillman RM, et al (1986). Relation of in vitro properties to tumorigenicity for a series of sublines of the human breast cancer cell line MCF-7. Cancer Res, 46, 6339-48.
  5. Castle VP, Dixit VM, Polverini PJ (1997). Thrombospondin-1 suppresses tumorigenesis and angiogenesis in serum- and anchorage-independent NIH 3T3 cells. Lab Invest, 77, 51-61.
  6. Chen J, Xie F, Zhang L, et al (2010). iASPP is over-expressed in human non-small cell lung cancer and regulates the proliferation of lung cancer cells through a p53 associated pathway. BMC Cancer, 10, 694. https://doi.org/10.1186/1471-2407-10-694
  7. Cui Y, Wu J, Zong M, et al (2009). Proteomic profiling in pancreatic cancer with and without lymph node metastasis. Int J Cancer, 124, 1614-21. https://doi.org/10.1002/ijc.24163
  8. Curto M, McClatchey AI (2004). Ezrin...a metastatic detERMinant? Cancer Cell, 5, 113-4. https://doi.org/10.1016/S1535-6108(04)00031-5
  9. Frisch SM, Francis H (1994). Disruption of epithelial cell-matrix interactions induces apoptosis. J Cell Biol, 124, 619-26. https://doi.org/10.1083/jcb.124.4.619
  10. Gautreau A, Louvard D, Arpin M (2002). ERM proteins and NF2 tumor suppressor: the Yin and Yang of cortical actin organization and cell growth signaling. Curr Opin Cell Biol, 14, 104-9. https://doi.org/10.1016/S0955-0674(01)00300-3
  11. Guo H, Ma Y, Zhang B, et al (2009). Pivotal advance: PKCzeta is required for migration of macrophages. J Leukoc Biol, 85, 911-8. https://doi.org/10.1189/jlb.0708429
  12. Gutierrez LZ, Suckow M, Lawler J, et al (2003). Thrombospondin 1--a regulator of adenoma growth and carcinoma progression in the APC(Min/+) mouse model. Carcinoqenesis, 24, 199-207. https://doi.org/10.1093/carcin/24.2.199
  13. Hoeflich KP, Ikura M (2004). Radixin: cytoskeletal adopter and signling protein. Int J Biochem Cell Biol, 36, 2131-6. https://doi.org/10.1016/j.biocel.2003.11.018
  14. Hoeflich KP, Tsukita S, Hicks L, et al (2003). Insights into a single rod-like helix in activated radixin required for membrane-cytoskeletal cross-linking. Biochemistry, 42, 11624-41.
  15. Jimenez B, Volpert OV, Crawford SE, et al (2000). Signals leading to apoptosis-dependent inhibition of neovascularization by thrombospondin-1. Nat Med, 6, 41-8. https://doi.org/10.1038/71517
  16. Khanna C, Wan X, Bose S, et al (2004). The membranecytoskeleton linker ezrin is necessary for osteosarcoma metastasis. Nat Med, 10, 182-6. https://doi.org/10.1038/nm982
  17. Kikuchi S, Hata M, Fukumoto K, et al (2002). Radixin deficiency causes conjugated hyperbilirubinemia with loss of Mrp2 from bile canalicular membranes. Nat Genet, 31, 320-5. https://doi.org/10.1038/ng905
  18. Kubo Y, Yoshii H, Kamiyama H, et al (2008). Ezrin, Radixin, and Moesin (ERM) proteins function as pleiotropic regulators of human immunodeficiency virus type 1 infection. Virology, 375, 130-40. https://doi.org/10.1016/j.virol.2008.01.047
  19. Lawler J, Miao WM, Duquette M, et al (2001). Thrombospondin-1 gene expression affects survival and tumor spectrum of p53- deficient mice. Am J Pathol, 159, 1949-56. https://doi.org/10.1016/S0002-9440(10)63042-8
  20. McClatchey AI (2003). Merlin and ERM proteins: unappreciated roles in cancer development? Nat Rev Cancer, 3, 877-83. https://doi.org/10.1038/nrc1213
  21. Meng Y, Lu Z, Yu S, et al (2010). Ezrin promotes invasion and metastasis of pancreatic cancer cells. J Transl Med, 8, 61. https://doi.org/10.1186/1479-5876-8-61
  22. Moon Y, Bottone FG Jr, McEntee MF, et al (2005). Suppression of tumor cell invasion by cyclooxygenase inhibitors is mediated by thrombospondin-1 via the early growth response gene Egr-1. Mol Cancer Ther, 4, 1551-8. https://doi.org/10.1158/1535-7163.MCT-05-0213
  23. Ou-Yang M, Liu HR, Zhang Y, et al (2011). ERM stable knockdown by siRNA reduced in vitro migration and invasion of human SGC-7901 cells. Biochimie, 93, 954-61. https://doi.org/10.1016/j.biochi.2011.01.017
  24. Paqlini G, Kunda P, Quiroqa S, et al (1998). Suppression of radixin and moesin alters growth cone morphology, motility, and process formation in primary cultured neurons. J Cell Biol, 143, 443-55. https://doi.org/10.1083/jcb.143.2.443
  25. Ramoni C, Luciani F, Spadaro F, et al (2002). Differential expression and distribution of ezrin, radixin and moesin in human natural killer cells. Eur J Immunol, 32, 3059-65. https://doi.org/10.1002/1521-4141(200211)32:11<3059::AID-IMMU3059>3.0.CO;2-3
  26. Revillion F, Lhotellier V, Hornez L, et al (2008). Real-time reverse-transcription PCR to quantify a panel of 19 genes in breast cancer: relationships with sentinel lymph node invasion. Int J Biol Markers, 23, 10-7. https://doi.org/10.1177/172460080802300102
  27. Singh S, Sadanandm A, Varney ML, et al (2010). Small Interfering RNA-mediated CXCR1 or CXCR2 Knock- DownInhibits Melanoma Tumor Growth and Invasion. Int J Cancer, 126, 328-36. https://doi.org/10.1002/ijc.24714
  28. Stemmler MP (2008). Cadherins in development and cancer. Mol Biosyst, 4, 835-50. https://doi.org/10.1039/b719215k
  29. Suda J, Zhu L, Karvar S (2011). Phosphorylation of radixin regulates cell polarity and Mrp-2 distribution in hepatocytes. Am J Physiol Cell Physiol, 300, C416-24. https://doi.org/10.1152/ajpcell.00467.2010
  30. Takeichi M (1993). Cadherins in cancer: implications for invasion and metastasis. Curr Opin Cell Biol, 5, 806-11. https://doi.org/10.1016/0955-0674(93)90029-P
  31. Tsukita S, Yonemura S (1999). Cortical actin organization: lessons from ERM (ezrin/radixin/moesin) proteins. J Biol Chem, 274, 34507-10. https://doi.org/10.1074/jbc.274.49.34507
  32. Turunen O, Wahlstrom T, Vaheri A (1994). Ezrin has a COOHterminal actin-binding site that is conserved in the ezrin protein family. J Cell Biol, 126, 1445-53. https://doi.org/10.1083/jcb.126.6.1445
  33. Van Roy F, Berx G (2008). The cell-cell adhesion molecule E-cadherin. Cell Mol Life Sci, 65, 3756-88. https://doi.org/10.1007/s00018-008-8281-1

Cited by

  1. Radixin Knockdown by RNA Interference Suppresses Human Glioblastoma Cell Growth in Vitro and in Vivo vol.15, pp.22, 2014, https://doi.org/10.7314/APJCP.2014.15.22.9805
  2. Prognostic Factors Influencing Clinical Outcomes of Malignant Glioblastoma Multiforme: Clinical, Immunophenotypic, and Fluorescence in Situ Hybridization Findings for 1p19q in 816 Chinese Cases vol.16, pp.3, 2015, https://doi.org/10.7314/APJCP.2015.16.3.971
  3. A glimpse of the ERM proteins vol.23, pp.1, 2016, https://doi.org/10.1186/s12929-016-0246-3