Imaging analysis of EGFR mutated cancer cells using peptide nucleic acid (PNA)-DNA probes

Hajime Shigeto, Takashi Ohtsuki, Akira Iizuka, Yasuto Akiyama, Shohei Yamamura

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Lung cancer cells harbor various gene mutations in the mRNA sequence of the Epidermal Growth Factor Receptor (EGFR), especially the mutations of exon19del E746-A750, T790M, and L858R. This results in cancer progression and resistance to anticancer drugs (tyrosine kinase inhibitor; TKI). Therefore, the imaging analysis of EGFR mutations is required for the treatment planning for non-small cell lung cancers. This study focused on the imaging analysis of a single nucleotide substitute in EGFR mutated cancer cells. We developed three novel peptide nucleic acid (PNA)-DNA probes for recognizing and detecting the following three gene mutations in EGFR gene mutations. The PNA-DNA probes consist of fluorescein isothiocyanate (FITC) conjugated PNA as a detection probe and Dabcyl conjugated DNA as a quencher probe. The PNA-DNA probes were used to validate the feasibility for detecting three EGFR mutated sequences: exon19del E746-A750, T790M, and L858R. The three probes emitted fluorescent dose-dependent signals against three target DNA and RNA. Using the three PNA-DNA probes, we succeeded in distinguishing three kinds of lung-cancer cell lines (H1975, PC-9, and A549) which have different EGFR mutations by the fluorescence in situ hybridization (FISH) method.

Original languageEnglish
Pages (from-to)4613-4621
Number of pages9
JournalAnalyst
Volume144
Issue number15
DOIs
Publication statusPublished - Aug 7 2019

Fingerprint

Nucleic Acid Probes
Peptide Nucleic Acids
DNA Probes
nucleic acid
Epidermal Growth Factor Receptor
peptide
cancer
DNA
Cells
probe
mutation
Imaging techniques
Mutation
Neoplasms
Genes
Lung Neoplasms
gene
erbB-1 Genes
Ports and harbors
Fluorescein

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy
  • Electrochemistry

Cite this

Imaging analysis of EGFR mutated cancer cells using peptide nucleic acid (PNA)-DNA probes. / Shigeto, Hajime; Ohtsuki, Takashi; Iizuka, Akira; Akiyama, Yasuto; Yamamura, Shohei.

In: Analyst, Vol. 144, No. 15, 07.08.2019, p. 4613-4621.

Research output: Contribution to journalArticle

Shigeto, Hajime ; Ohtsuki, Takashi ; Iizuka, Akira ; Akiyama, Yasuto ; Yamamura, Shohei. / Imaging analysis of EGFR mutated cancer cells using peptide nucleic acid (PNA)-DNA probes. In: Analyst. 2019 ; Vol. 144, No. 15. pp. 4613-4621.
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