Silencing endothelin-3 expression attenuates the malignant behaviors of human melanoma cells by regulating SPARC levels

J Huazhong Univ Sci Technolog Med Sci. 2013 Aug;33(4):581-586. doi: 10.1007/s11596-013-1162-3. Epub 2013 Aug 1.

Abstract

Endothelin-3 (ET-3) is aberrantly expressed in both metastatic melanoma tissues and cultured melanoma cells. Our previous work showed that ET-3 could promote survival of metastatic melanoma cells via its altered expression. In this study, we investigated the mechanisms responsible for these gene-induced phenotypes in melanoma cells. An ET-3 gene sequence-specific shRNA vector pLVTHM-ET3-RNAi was constructed and transfected into human malignant melanoma cells A375 and MMRU, and the resultant molecular events and cellular changes were examined. As compared with the empty-vector group, cell proliferation was slowed down, and the growth inhibition rates were 38.9% in A375 cells and 38.4% in MMRU cells after transfection. In addition, cell invasion capability was also inhibited, with a reduction of 62.2% in A375 cells and 54.3% in MMRU cells. The percentage of apoptotic cells was found to increase. Meanwhile, in both cell lines, secreted protein acidic and rich in cysteine (SPARC) levels were down-regulated together with inhibition of its upstream signaling molecule, NF-κB. Thus, the current results suggested that down-regulated expression of ET3 attenuates the malignant behaviors of human melanoma cells partially by decreasing the expression of SPARC and NF-κB.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Endothelin-3 / genetics*
  • Gene Silencing
  • Humans
  • Melanoma / genetics*
  • Melanoma / pathology*
  • Osteonectin / genetics*

Substances

  • Endothelin-3
  • Osteonectin
  • SPARC protein, human