Effect Of Metallothionein Silencing On Metastasis Gene In The Breast Cancer Cell

Objectives Breast cancer is a common malignant tumor. To date, many genes associated with the development and transfer of breast cancer have been identified. This study intends to observe the proliferation of breast cancer cells and some genes expression associated with breast cancer metastasis in MT2 A gene silence MCF-7 breast cancer cell.Methods The sequence characteristics of metallothionein MT2 A gene was analyzed using bioinformatics, and then the vector sh RNA-MT2 A was designed and constructed. The breast cancer cell lines were obtained via transferring vector sh RNA-MT2 A into MCF-7 breast cancer cell. The proliferation and adhesion matrix changes of MT2 A gene silencing breast cancer cells were observed by RT-PCR and western blot experiments. The expression changes of FGF-1, Twist, transferrin, AOP-1 and peroxiredoxin 6 were analyzed by by RT-PCR and western blot experiments.Results 1 A sh RNA-MT2 A vector from designing and constructing three sh RNA-MT2 A vector were finally determined for subsequent research through the experiment.2 The MT2 A gene silencing breast cancer cells were obtained by liposome transfection technique, and identified by semi-quantitative PCR and western blot experiments.3 The proliferation of MT2 A gene silencing breast cancer cells reduced compared with wild cell lines; The adhesion matrix of MT2 A gene silencing breast cancer cells improved compared with wild cell lines.4 The real time quantitative PCR results showed that the expression level of FGF-1 and Twist in MT2 A gene silencing breast cancer cells haven’t significant change.5 The expression level of transferrin in MT2 A gene silencing breast cancer cells was significant upregulated in MT2 A gene silencing breast cancer cells.6 The study confirmed that the m RNA level of AOP-1 and peroxiredoxin 6 were higher than that in wild cells.7 Western blot experiments also showed that the protein level of FGF-1 and Twists in MT2 A gene silencing breast cancer cell did not significantly changed, but the protein level of transferring, AOP-1 and peroxiredoxin 6 in MT2 A gene silencing breast cancer cell were higher than that in wild cells.8 The overexpression of MT2 A and beta-domain in breast cancer cell did not influence cell proliferation, however, the overexpression of beta-domain in breast cancer cell up-regulate the expression of cytochrome c oxidase subunit 6B2.Conclusions This study confirmed that the MT2 A gene silencing can inhibit the proliferation of breast cancer cell, and enhance its matrix adhesion ability. MT2 A gene silencing also can upregulate the expression of transferring, cytochrome c oxidase subunit 6B2, AOP-1 and peroxiredoxin 6, because the controlling metal ions metabolism and antioxidant are the main function of MT.