Expression of hPNAS-4 radiosensitizes Lewis lung cancer

Int J Radiat Oncol Biol Phys. 2012 Nov 15;84(4):e533-40. doi: 10.1016/j.ijrobp.2012.06.028. Epub 2012 Jul 24.

Abstract

Purpose: This study aimed to transfer the hPNAS-4 gene, a novel apoptosis-related human gene, into Lewis lung cancer (LL2) and observe its radiosensitive effect on radiation therapy in vitro and in vivo.

Methods and materials: The hPNAS-4 gene was transfected into LL2 cells, and its expression was detected via western blot. Colony formation assay and flow cytometry were used to detect the growth and apoptosis of cells treated with irradiation/PNAS-4 in vitro. The hPNAS-4 gene was transferred into LL2-bearing mice through tail vein injection of the liposome/gene complex. The tumor volumes were recorded after radiation therapy. Proliferating cell nuclear antigen (PCNA) immunohistochemistry staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay were performed to detect the tumor cell growth and apoptosis in vivo.

Results: The hPNAS-4 gene was successfully transferred into LL2 cells and tumor tissue, and its overexpressions were confirmed via western blot analysis. Compared with the control, empty plasmid, hPNAS-4, radiation, and empty plasmid plus radiation groups, the hPNAS-4 plus radiation group more significantly inhibited growth and enhanced apoptosis of LL2 cells in vitro and in vivo (P<.05).

Conclusions: The hPNAS-4 gene was successfully transferred into LL2 cells and tumor tissue and was expressed in both LL2 cell and tumor tissue. The hPNAS-4 gene therapy significantly enhanced growth inhibition and apoptosis of LL2 tumor cells by radiation therapy in vitro and in vivo. Therefore, it may be a potential radiosensitive treatment of radiation therapy for lung cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis
  • Apoptosis Regulatory Proteins / genetics
  • Apoptosis Regulatory Proteins / metabolism*
  • Carbon-Nitrogen Lyases
  • Carcinoma, Lewis Lung / metabolism*
  • Carcinoma, Lewis Lung / pathology
  • Carcinoma, Lewis Lung / therapy*
  • Cell Line, Tumor
  • Female
  • Flow Cytometry
  • Humans
  • In Situ Nick-End Labeling / methods
  • Lung Neoplasms / metabolism*
  • Lung Neoplasms / pathology
  • Lung Neoplasms / therapy*
  • Mice
  • Mice, Inbred C57BL
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism*
  • RNA, Small Interfering / pharmacology
  • Random Allocation
  • Transfection / methods
  • Tumor Burden
  • Tumor Stem Cell Assay

Substances

  • Apoptosis Regulatory Proteins
  • Neoplasm Proteins
  • RNA, Small Interfering
  • Carbon-Nitrogen Lyases
  • DESI2 protein, human