The Establishment and Application of the Method Detecting hMAM Gene Expression of Breast Cancer Cell Lines in Peripheral Blood / 医学研究杂志

ABSTRACT

Objective To develop a method detecting hMAM gene expression of breast cancer cell lines in peripheral blood.Methods The hMAM gene was cloned and confirmed by agarose gel electrophoresis and sequence analysise,the standard substance of real-time quantitative polymerase chain reaction (FQ-PCR )was prepared. The breast cancer cell lines MDA-MB453、MCF7 were cultured and the blood sample model was made.The hMAM expression of breast cancer cell lines blood model and 10 cases of breast cancer patient peripheral blood were detected by RT-PCR and FQ-PCR,GAPDH gene expression as contrast.Results The production of PCR was cloned and testified by sequence.The correlation coefficient(r) of standard curve line of FQ-PCR was -1.0. The x?s and CV were 718.9?120.5 and 16.7% respectively.The hMAM expression of one breast cancer cell lines MDA-MB453 in 104 blood cells could be detected, but not detected in the breast cancer cell lines MCF7. The hMAM expression was detected in 3 of 5 patients with sentinel nodes and in 1 of 5 patients with non-sentinel nodes.Conclusions It might be a good method for detecting breast cancer cell micrometastased in peripheral blood. The diffrernce of hMAM gene expression correlates with the type of breast cancer cell lines.