Inhibition of CD146 gene expression via RNA interference reduces in vitro perineural invasion on ACC-M cell

J Oral Pathol Med. 2009 Feb;38(2):198-205. doi: 10.1111/j.1600-0714.2008.00706.x.

Abstract

Background: Overexpression of melanoma cell adhesion molecule (MCAM or CD146), a novel member of the immunoglobulin gene superfamily, promotes invasion, metastasis, growth and survival of malignant cells.

Materials and methods: Here, we used a human U6 promoter-driven DNA template approach to induce short hairpin RNA (shRNA)-triggered RNA interference to block CD146 gene expression in the human high-metastatic adenoid cystic carcinoma (ACC) cell line ACC-M. reverse transcription-polymerase chain reaction, Western blot, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay, flow cytometry analysis and in vitro perineural invasion (PNI) assay were used to investigate the effects of CD146 on cell growth, cell cycle distribution and cell PNI activity in ACC-M cells.

Results: We have demonstrated that different shRNAs can specifically knockdown the transcription of CD146, resulting in the inhibition of cell proliferation, and G(0)/G(1) to S phase arrest in ACC-M cells. Knockdown of CD146 by shRNA resulted in decrease of the ACC-M PNI activity in vitro.

Conclusion: These findings suggested that CD146 was an ACC-related gene and CD146 may play an important role in the carcinogenesis, progression and PNI activity of ACC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • CD146 Antigen / biosynthesis*
  • CD146 Antigen / genetics
  • Carcinoma, Adenoid Cystic / genetics
  • Carcinoma, Adenoid Cystic / pathology*
  • Cell Cycle Proteins / metabolism
  • Cell Line, Tumor
  • Down-Regulation
  • Gene Expression
  • Gene Expression Regulation, Neoplastic
  • Gene Knockdown Techniques
  • Humans
  • Neoplasm Invasiveness / pathology*
  • Peripheral Nerves / pathology
  • RNA Interference*
  • RNA, Small Interfering / physiology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transfection

Substances

  • CD146 Antigen
  • Cell Cycle Proteins
  • RNA, Small Interfering