[Analyzed the molecular interaction network of tumor suppressor gene 14-3-3 sigma in lung cancer cell based on stable isotope labeling by amino acids in cell culture technology]

Zhonghua Yu Fang Yi Xue Za Zhi. 2013 Aug;47(8):752-6.
[Article in Chinese]

Abstract

Objective: To analysis the molecular interaction network of 14-3-3 sigma in non small cell lung cancer (NSCLC) cells.

Methods: Established stable over-expressed 14-3-3 sigma protein PG cells, MTT assay was used to assess the growth rate of PG cells. Though stable isotope labeling by amino acids in cell culture (SILAC) and Mass spectrometry (MS) technology, to identify difference expressed proteins caused by over expressed 14-3-3 sigma. The protein expressed >2 or <0.5 times was termed as the differential protein. By searching Human protein reference database (HPRD) and Kyoto encyclopedia of genes and genomes (KEGG), established the molecular interaction network of tumor suppressor gene 14-3-3 sigma.

Results: The growth rate of over-expressed 14-3-3 sigma PG cell was obviously slower down compared to vector PG cells. A database including 147 differential protein was established. And a molecular interaction network of 14-3-3 sigma containing 26 protein was constructed.In this network, the expression of CSNK2A1 (casein kinase II subunit alpha), involved in numerous cellular processes, such as cell cycle progression, apoptosis and transcription, was the most significantly increased. A DNA repair protein, MEN1 (Menin) which functions as a transcriptional regulator was the most significantly decreased.

Conclusion: After stable transfected with 14-3-3 sigma gene, growth rate of PG cells was inhibited, the proteins associated with cell cycle, DNA damage repair mechanisms were significantly changed, and constructed the molecular interaction network.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 14-3-3 Proteins / genetics*
  • Amino Acids
  • Biomarkers, Tumor / genetics*
  • Carcinoma, Non-Small-Cell Lung / genetics
  • Cell Cycle
  • Cell Line, Tumor
  • Cell Proliferation
  • Exoribonucleases / genetics*
  • Humans
  • Isotope Labeling / methods
  • Lung Neoplasms / genetics*
  • Mass Spectrometry
  • Transfection

Substances

  • 14-3-3 Proteins
  • Amino Acids
  • Biomarkers, Tumor
  • Exoribonucleases
  • SFN protein, human