| BackgroundAs mentioned in the Canon of 《yellow emperor, nine stitches on paper》that theguests of four seasons and eight winds in channels and collaterals are for tumordisease. It puts forward the pathogenesis of tumor disease that the six exogenous evilsstayed invaded into main and collateral channels. And more breast tumor is due toliver meridian impassability, vital qi deficiency caused by the attacks. Breast cancer isa common malignant tumor of female ranked first, the American journal of clinicalcancer doctors, according to the latest statistics released in2011,230480cases offemale breast cancer in the United States, accounting for30%of the new female hairmalignant tumor Beijing, Shanghai, Tianjin and other big cities in our countryaccording to breast cancer is also a women of the most common malignant tumor inour country, and there is an upward trend in incidence of a disease, breast cancerdevelopment is directly related to endocrine peptide. Neuroendocrine peptides are thebody’s endocrine, paracrine or autocrine or living cells secrete active peptides, thereare more than350in the body, the mature peptides need nine protein shift enzyme(Pro-protein Convertases, PCs) synergy, to each other in time and space processing neuroendocrine peptide precursor protein (protein) in specific tissues and cells ofmature. Pro-protein Convertase2(PC2) gene is a cell’s normal, PC2enzyme mediatedthe processing system of present low levels in breast cancer, this change is closelyrelated to tumourgenesis, development and apoptosis. This study researches the stateof breast cancer cell proliferation in the pc2gene expression.ObjectiveStudy the influence of the regulating PC2expression in the tumorigenesis,development and metastasis of breast cancer cells for providing a new research area indiagnosis and treatment of breast cancer.MethodsRNA was extracted from rat pituitary tissue after RT-PCR, the recombinant vectorand screening means to obtain the target gene, and combined with agarose gelelectrophoresis restructuring pc2and7b2gene identification. Plasmid extraction kit (asmall amount) and ion-exchange column (large), extracted and purified therecombinant gene. The preparation of liposome-mediated gene was transferred intothe cancer cells. PC2protein expression was analysised by Western blot. Analysis ofthe role of the PC2gene on the proliferation of cancer cells by MTT assay andfluorescence immunoassay in vitro experiments. The pc2gene formulation wasevaluated of the proliferation of cancer cells by Ex Vivo and In vivo miceexperiments.ResultsIn this study, recombinant PC2and7b2gene was successfully constructed andusing liposome-mediated technology PC2gene transfected into cancer cells. Westernblot results showed that level of the PC2protein expression of the pc2and7b2genetransfected4T1cells was higher than the wild-type4T1cells which indicated that thegene was successfully transfected into cells. The in vitro inhibition of MTTexperiment results show that with the increase in the amount pc2and7b2gene transfection, the proliferation of cancer cells is suppressed. In immunofluorescenceexperiments, the nucleus of the cells which transfectd pc2and7b2gene break, whilethe wild-type cell nucleus remained circular. After injection of cancer cells transfectedgene in mice,7mice in the control group tumor, while the the pc2and7b2group ofmice grow only one tumor. Vivo gene therapy of tumor-bearing mice, the pc2and7b2group of tumors whose growth was inhibited, whereas the control group tumors togrow over time.ConclusionUp-regulation of pc2and7b2gene can inhibit the proliferation of breast cancercells and has the dose dependent relationship for the inhibition. |
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