Lentivirus-delivered ZEB-1 small interfering RNA inhibits lung adenocarcinoma cell growth in vitro and in vivo

J Cancer Res Clin Oncol. 2012 Aug;138(8):1329-38. doi: 10.1007/s00432-012-1206-2. Epub 2012 Apr 6.

Abstract

Purpose: To explore the relationship between the expression of ZEB1 gene and the proliferation ability of lung adenocarcinoma cells.

Methods: Immunohistochemistry, Western blot and Real-time PCR were used to detect the expression of ZEB1 gene in lung adenocarcinoma tissue and cell lines compared with adjacent noncancerous region and the human lung fibroblast cell HLF cells. The lentivirus RNA interference technique was used to knock down the expression of ZEB1 in lung adenocarcinoma A549 and H1299 cell lines. Cell cycle and cell apoptosis were measured by FCM assay. In vivo, four groups of 4-week-old nude mice were subcutaneously injected with the stably transfected (ZEB-si, scr-si) cells at a single site to investigate the effect of ZEB1-siRNA in the nude mice tumor growth. In situ apoptosis was detection by TUNEL assay.

Results: ZEB-1 was highly expressed in lung adenocarcinoma tissue and cell lines compared with adjacent noncancerous region and the human lung fibroblast cell HLF cells. ZEB1-siRNA could decrease lung adenocarcinoma cell proliferation by delaying S-phase entry and induce cell apoptosis, which led to the inhibition of the tumorigenicity of A549 and H1299 cell lines. Further investigation showed that injecting the ZEB1-siRNA cells into the nude mice could significantly decrease the tumor growth.

Conclusion: Knockdown of ZEB-1 expression by lentivirus-delivered siRNA may provide a novel therapeutic target for the treatment of lung cancer.

MeSH terms

  • Adenocarcinoma / genetics*
  • Adenocarcinoma / pathology
  • Adenocarcinoma / therapy
  • Animals
  • Blotting, Western
  • Cell Cycle / genetics
  • Cell Line, Tumor
  • Cell Proliferation*
  • Female
  • Gene Expression Regulation, Neoplastic
  • Homeodomain Proteins / genetics*
  • Homeodomain Proteins / metabolism
  • Humans
  • Immunohistochemistry
  • Lentivirus / genetics
  • Lung Neoplasms / genetics*
  • Lung Neoplasms / pathology
  • Lung Neoplasms / therapy
  • Male
  • Mice
  • Mice, Nude
  • Middle Aged
  • RNA Interference
  • RNA, Small Interfering / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • S Phase / genetics
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism
  • Tumor Burden / genetics
  • Xenograft Model Antitumor Assays / methods
  • Zinc Finger E-box-Binding Homeobox 1

Substances

  • Homeodomain Proteins
  • RNA, Small Interfering
  • Transcription Factors
  • ZEB1 protein, human
  • Zinc Finger E-box-Binding Homeobox 1