PMID- 31241068
OWN - NLM
STAT- MEDLINE
DCOM- 20191206
LR  - 20191217
IS  - 1364-5528 (Electronic)
IS  - 0003-2654 (Linking)
VI  - 144
IP  - 15
DP  - 2019 Aug 7
TI  - Imaging analysis of EGFR mutated cancer cells using peptide nucleic acid 
      (PNA)-DNA probes.
PG  - 4613-4621
LID - 10.1039/c9an00725c [doi]
AB  - 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.
FAU - Shigeto, Hajime
AU  - Shigeto H
AUID- ORCID: 0000-0003-0602-8736
AD  - Health Research Institute, National Institute of Advanced Industrial Science and 
      Technology (AIST), 2217-14 Hayashi-cho, Takamatsu, Kagawa 761-0395, Japan. 
      hajime.shigeto@aist.go.jp yamamura-s@aist.go.jp.
FAU - Ohtsuki, Takashi
AU  - Ohtsuki T
AUID- ORCID: 0000-0002-3183-5305
AD  - Department of Interdisciplinary Science and Engineering in Health Systems, 
      Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan.
FAU - Iizuka, Akira
AU  - Iizuka A
AD  - Immunotherapy Division, Shizuoka Cancer Center Research Institute, Shizuoka 
      411-8777, Japan.
FAU - Akiyama, Yasuto
AU  - Akiyama Y
AD  - Immunotherapy Division, Shizuoka Cancer Center Research Institute, Shizuoka 
      411-8777, Japan.
FAU - Yamamura, Shohei
AU  - Yamamura S
AUID- ORCID: 0000-0002-4971-4560
AD  - Health Research Institute, National Institute of Advanced Industrial Science and 
      Technology (AIST), 2217-14 Hayashi-cho, Takamatsu, Kagawa 761-0395, Japan. 
      hajime.shigeto@aist.go.jp yamamura-s@aist.go.jp.
LA  - eng
PT  - Journal Article
DEP - 20190626
PL  - England
TA  - Analyst
JT  - The Analyst
JID - 0372652
RN  - 0 (DNA Probes)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Peptide Nucleic Acids)
RN  - 0 (RNA, Messenger)
RN  - 6268-49-1 (4-(4-dimethylaminophenylazo)benzoic acid)
RN  - 9007-49-2 (DNA)
RN  - A49L8E13FD (p-Dimethylaminoazobenzene)
RN  - EC 2.7.10.1 (EGFR protein, human)
RN  - EC 2.7.10.1 (ErbB Receptors)
RN  - I223NX31W9 (Fluorescein-5-isothiocyanate)
SB  - IM
MH  - Cell Line, Tumor
MH  - DNA/*genetics
MH  - DNA Probes/*chemistry/genetics
MH  - ErbB Receptors/genetics
MH  - Fluorescein-5-isothiocyanate/chemistry
MH  - Fluorescent Dyes/chemistry
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Lung Neoplasms/classification/genetics
MH  - Nucleic Acid Hybridization
MH  - Peptide Nucleic Acids/*chemistry/genetics
MH  - Point Mutation
MH  - RNA, Messenger/*genetics
MH  - p-Dimethylaminoazobenzene/analogs & derivatives/chemistry
EDAT- 2019/06/27 06:00
MHDA- 2019/12/18 06:00
CRDT- 2019/06/27 06:00
PHST- 2019/06/27 06:00 [pubmed]
PHST- 2019/12/18 06:00 [medline]
PHST- 2019/06/27 06:00 [entrez]
AID - 10.1039/c9an00725c [doi]
PST - ppublish
SO  - Analyst. 2019 Aug 7;144(15):4613-4621. doi: 10.1039/c9an00725c. Epub 2019 Jun 26.