sybr-green-i and Carcinoma--Ductal--Breast

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.. 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.. 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.. 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.

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

Growing evidence suggests microRNAs (miRNAs) have an important role in tumorigenesis. MicroRNA-21 (miR-21) is up-regulated in many malignant tumors, including breast cancer. Its association with clinicopathologic features and expression of PTEN (phosphatase and tensin homolog deleted on chromosome 10), one of its target genes, in breast cancer has not been reported systematically. To further determine the potential involvement of miR-21 in breast cancer, we have evaluated the expression level of miR-21 by stem-loop real-time RT-PCR based on SYBR-Green I in human invasive ductal carcinoma of the breast, and we have correlated the results with clinicopathologic features and PTEN protein expression. Matched non-tumor and tumor tissues of 40 human invasive ductal carcinoma of the breast were analyzed for miR-21 expression by stem-loop real-time RT-PCR based on SYBR-Green I. Immunohistochemistry (IHC) was used to estimate PTEN expression in tumor tissue. The expression levels of miR-21 were correlated with PTEN and commonly used clinicopathologic features of breast cancer. The stem-loop real-time RT-PCR based on SYBR-Green I was sensitive and specific enough to detect miR-21. Expression levels of miR-21 were significantly higher in tumor tissues than the levels in matched non-tumor tissues (P=0.000). Expression of miR-21 was negatively correlated with expression of PTEN (P=0.013). Up-regulated miR-21 expression was associated with lymph node positivity (P=0.01), higher proliferation index (ki67>10%) (P=0.03) and advanced breast cancer TNM clinical stage (P=0.021). These findings suggest that PTEN is possibly one of the targets of miR-21 in breast cancer and high expression of mir-21 indicates a more aggressive phenotype.

Topics: Benzothiazoles; Breast Neoplasms; Carcinoma, Ductal, Breast; Diamines; Female; Humans; Immunohistochemistry; MicroRNAs; Neoplasm Staging; Organic Chemicals; PTEN Phosphohydrolase; Quinolines; Reverse Transcriptase Polymerase Chain Reaction