Detection of EGFR mutation in supernatant, cell pellets of pleural effusion and tumor tissues from non-small cell lung cancer patients by high resolution melting analysis and sequencing

Int J Clin Exp Pathol. 2014 Dec 1;7(12):8813-22. eCollection 2014.

Abstract

To determine epidermal growth factor receptor (EGFR) mutation in advanced non-small cell lung cancer (NSCLC) patients and compare the detection efficiency between different sample resources, both high resolution melting (HRM) analysis and direct sequencing method were used to analyze 36 pleural effusion samples and 22 matched biopsy tumor tissues collected from NSCLC patients. For each pleural effusion sample, the supernatant and the cell pellets were examined separately. Among all the 36 cases of pleural effusion samples, 18 mutations of EGFR were found in cell-free supernatant while 13 mutations were found in the cell pellets as detected by HRM analysis. In the 22 matched samples, 13 cases of EGFR mutations were identified in paraffin-embedded biopsy tissue samples, 12 cases in the cell-free supernatant and 9 cases in the cell pellets of pleural effusion. EGFR mutations in 15 cases out of the total 36 pleural effusion samples detected by direct sequencing were also identified by HRM analysis, giving 100% efficiency for HRM method. The results established the important role of HRM as a reliable and efficient method to determine EGFR mutation status and indicated the feasibility of using pleural effusion in replacement of biopsy tissues in particular clinical cases. Furthermore, the cell-free supernatant of pleural effusion might be a better resource for mutation detection than cell pellets.

Keywords: EGFR; HRM; NSCLC; pleural effusion; supernatant.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Base Sequence
  • Carcinoma, Non-Small-Cell Lung / genetics*
  • DNA Mutational Analysis / methods*
  • DNA, Neoplasm / analysis*
  • ErbB Receptors / analysis*
  • Female
  • Humans
  • Lung Neoplasms / genetics*
  • Male
  • Middle Aged
  • Mutation*
  • Pleural Effusion, Malignant / genetics
  • Polymerase Chain Reaction

Substances

  • DNA, Neoplasm
  • EGFR protein, human
  • ErbB Receptors