Detection of CCND1 , C-MYC , and FGFR1 amplification using modified SYBR Green qPCR and FISH in breast cancer

Turk J Med Sci. 2018 Aug 16;48(4):759-767. doi: 10.3906/sag-1710-93.

Abstract

Background/aim: The aims of this study were to detect CCND1 , C-MYC , and FGFR1 amplification using qPCR, confirmation with FISH, and to further assess their clinicopathological relevance.

Materials and methods: Thirty-five breast tumor samples were analyzed for amplification of the selected genes using modified SYBR Green qPCR. The accuracy of the qPCR was assessed by FISH as a gold-standard method.

Results: CCND1 , C-MYC , and FGFR1 amplifications were observed in 34.28%, 28.57%, and 17.14% of the 35 samples, respectively. qPCR results were significantly confirmed by FISH and qPCR and FISH showed excellent correlation (P = 0.000). CCND1 amplification with tumor stage (P = 0.044), positive metastatic status (P = 0.042), positive family history (P = 0.042), and C-MYC status (P = 0.005); C-MYC amplification with tumor size (P = 0.021), tumor grade (P = 0.018), tumor stage (P = 0.032), and FGFR1 status (P < 0.000); and FGFR1 amplification with tumor size (P = 0.041) and positive ER status (P = 0.042) were statistically associated.

Conclusion: Our findings revealed that the applied qPCR approach could precisely quantify the relative gene copy number. More studies with a larger sample size are suggested to confirm the clinicopathological value of CCND1 , C-MYC , and FGFR1 amplification.

Keywords: Breast cancer; fluorescent in situ hybridization; gene amplification; gene copy number; real-time polymerase chain reaction.

Publication types

  • Validation Study

MeSH terms

  • Adolescent
  • Adult
  • Benzothiazoles
  • Breast Neoplasms / genetics*
  • Breast Neoplasms / pathology
  • Carcinoma, Ductal, Breast / genetics*
  • Cyclin D1 / genetics*
  • DNA-Binding Proteins / genetics*
  • Diamines
  • Female
  • Gene Amplification*
  • Gene Dosage
  • Humans
  • In Situ Hybridization, Fluorescence / methods
  • Middle Aged
  • Neoplasm Grading
  • Neoplasm Staging
  • Organic Chemicals
  • Quinolines
  • Real-Time Polymerase Chain Reaction / methods*
  • Receptor, Fibroblast Growth Factor, Type 1 / genetics*
  • Reproducibility of Results
  • Transcription Factors / genetics*
  • Young Adult

Substances

  • Benzothiazoles
  • CCND1 protein, human
  • DNA-Binding Proteins
  • Diamines
  • MYCBP protein, human
  • Organic Chemicals
  • Quinolines
  • Transcription Factors
  • Cyclin D1
  • SYBR Green I
  • Receptor, Fibroblast Growth Factor, Type 1