PMID- 26765781
OWN - NLM
STAT- MEDLINE
DCOM- 20160711
LR  - 20181113
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 11
IP  - 1
DP  - 2016
TI  - Detection of MET Gene Copy Number in Cancer Samples Using the Droplet Digital PCR 
      Method.
PG  - e0146784
LID - 10.1371/journal.pone.0146784 [doi]
LID - e0146784
AB  - PURPOSE: The analysis of MET gene copy number (CN) has been considered to be a 
      potential biomarker to predict the response to MET-targeted therapies in various 
      cancers. However, the current standard methods to determine MET CN are SNP 6.0 in 
      the genomic DNA of cancer cell lines and fluorescence in situ hybridization 
      (FISH) in tumor models, respectively, which are costly and require advanced 
      technical skills and result in relatively subjective judgments. Therefore, we 
      employed a novel method, droplet digital PCR (ddPCR), to determine the MET gene 
      copy number with high accuracy and precision. METHODS: The genomic DNA of cancer 
      cell lines or tumor models were tested and compared with the MET gene CN and 
      MET/CEN-7 ratio determined by SNP 6.0 and FISH, respectively. RESULTS: In cell 
      lines, the linear association of the MET CN detected by ddPCR and SNP 6.0 is 
      strong (Pearson correlation = 0.867). In tumor models, the MET CN detected by 
      ddPCR was significantly different between the MET gene amplification and 
      non-amplification groups according to FISH (mean: 15.4 vs 2.1; P = 0.044). Given 
      that MET gene amplification is defined as MET CN >5.5 by ddPCR, the concordance 
      rate between ddPCR and FISH was 98.0%, and Cohen's kappa coefficient was 0.760 
      (95% CI, 0.498-1.000; P <0.001). CONCLUSIONS: The results demonstrated that the 
      ddPCR method has the potential to quantify the MET gene copy number with high 
      precision and accuracy as compared with the results from SNP 6.0 and FISH in 
      cancer cell lines and tumor samples, respectively.
FAU - Zhang, Yanni
AU  - Zhang Y
AD  - Amgen Biopharmaceutical Research & Development (Shanghai) Co., Ltd, Shanghai, 
      China.
FAU - Tang, En-Tzu
AU  - Tang ET
AD  - Amgen Biopharmaceutical Research & Development (Shanghai) Co., Ltd, Shanghai, 
      China.
FAU - Du, Zhiqiang
AU  - Du Z
AD  - Amgen Biopharmaceutical Research & Development (Shanghai) Co., Ltd, Shanghai, 
      China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20160114
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Biomarkers, Tumor)
RN  - EC 2.7.10.1 (MET protein, human)
RN  - EC 2.7.10.1 (Proto-Oncogene Proteins c-met)
SB  - IM
MH  - Biomarkers, Tumor
MH  - Cell Line, Tumor
MH  - *DNA Copy Number Variations
MH  - Gene Amplification
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Neoplasms/drug therapy/*genetics/pathology
MH  - Polymerase Chain Reaction/methods
MH  - Polymorphism, Single Nucleotide
MH  - Proto-Oncogene Proteins c-met/*genetics
PMC - PMC4713204
COIS- Competing Interests: All authors are employees of Amgen Inc. This commercial 
      affiliation did not alter the authors' adherence to PLOS ONE policies on sharing 
      data and materials.
EDAT- 2016/01/15 06:00
MHDA- 2016/07/12 06:00
PMCR- 2016/01/14
CRDT- 2016/01/15 06:00
PHST- 2015/09/18 00:00 [received]
PHST- 2015/12/22 00:00 [accepted]
PHST- 2016/01/15 06:00 [entrez]
PHST- 2016/01/15 06:00 [pubmed]
PHST- 2016/07/12 06:00 [medline]
PHST- 2016/01/14 00:00 [pmc-release]
AID - PONE-D-15-41268 [pii]
AID - 10.1371/journal.pone.0146784 [doi]
PST - epublish
SO  - PLoS One. 2016 Jan 14;11(1):e0146784. doi: 10.1371/journal.pone.0146784. 
      eCollection 2016.