Development and primary application of a real-time quantitative polymerase chain reaction for the detection of human breast cancer related novel gene-Metadherin expression

Abstract

Objective To establish a quantitative SYBR GreenⅠreal-time PCR method for detection of Metadherin (MTDH) gene expression level in breast cancer, and to investigate the relationship between MTDH mRNA expression level and the clinicopathological characters. Methods Real-time PCR was employed to determine the expression level of MTDH mRNA in peripheral blood of 80 specimens of breast cancer patients and normal females. Results In the 80 specimens, MTDH mRNA was positively expressed in the peripheral blood of 61 cases of breast cancer patients while negatively expressed in the 19 cases of normal females. Among the 61 breast cancer patients, MTDH mRNA showed high expression in 34 cases, accounting for 55.7%, and showed low expression in 27 cases, accounting for 44.3%. Both of the differences in expression rate has statistic significance (Ratio = 2.02 ± 0.81, P<0.05). MTDH expression relative to GAPDH expression in the peripheral blood of breast cancer patients was 1.15 ± 0.36. MTDH mRNA expression has no connection with age, estrogen and progesterone receptors , as well as HER-2.(P＞0.05). There is statistical difference for MTDH mRNA expression level between the group with lymph node metastasis and the group without lymph node metastasis (Ratio=2.02 ± 0.81,P<0.05). MTDH mRNA expression level changed between clinicopathological stage I, II and III, IV. Conclusion The established SYBR Green Ⅰ quantitative real-time PCR method can successfully detect the expression level of MTDH mRNA, which may be closely related to the occurrence and development of breast cancer. Key words: Breast cancer; MTDH; Real-time fluorescence quantitative PCR