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

Abstract

Background/aim: The aims of this study were to detect CCND1, C-MYC, and FGFR1 amplification using qPCR, confirmation withFISH, 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 SYBRGreen 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 amplificationwith 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); andFGFR1 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. Morestudies with a larger sample size are suggested to confirm the clinicopathological value of CCND1, C-MYC, and FGFR1 amplification.