Effects Of ADAM17-siRNA On The Proliferation Of MCF-7 Breast Cancer Cell

ObjectiveBreast tumor is the most common malignant tumor in female, its incidence is increasing in recent years. For now, its pathogenic mechanism is not clear but it is considered that it results from multiple actions among many factors in coordination with many genes and protein high expression in breast cancer, including oncogene active and antioncogene defective. In recent years, More and more researchers have attracted much attention in the field of genetic changes and function analysis in the development and prognosis of breast cancer. It has been recently discovered that A Disintegrin And Metalloproteinase is of the same category of multifunctional cell membranecell protein family, which is made up of 800 to 1200 amino acid. ADAM, as a new target, may play a valuable role in tumor target therapy by affecting transduction of EGRF signal. ADAM17, TACE, is one of the members of ADAMs, which has been observed high expression in multiple tumors, and played a significant role in tumor genesis and development by regulating cell cycle. Our published reports further confirmed that ADAM17 protein was observed low expression in fibroadenoma, but high expression in breast cancer tissue.RNAi (RNA interference) technology has been commonly used in the approach of silencing gene expression recently. Compared with antisense oligonucleotides, RNA interference technology has following advantages: higher efficiency, better specificity, lower toxicity, quicker effect, as well as more convenient manipulation. So it will gradually substitute traditional antisense technology in gene block.The study was designed to investigate the effect of ADAM17-si- RNA on the growth and proliferation of MCF-7 breast cancer cell line by making ADAM17 gene silence with siRNA. The research is expected to reveal the relationship between ADAM17 and cell cycle regulation and establish the theory basis for target gene therapy.MethodsRNAi technology was used to observe its inhibitory effect on the growth of human breast cancer MCF-7 cells. siRNA targeting ADAM17 was transfected into MCF-7 cells mediated by oligofectamine. ADAM17 mRNA expression were detected by RT-PCR after transfection. The phenotypic change of MCF-7 cells including proliferation ability and cell cycle after RNAi trasfection was studied by MTT assay and flow cytometry.ResultsRT-PCR revealed significantly lowered ADAM17 expression at mRNA level in transfected MCF-7 cells. MCF-7 cells exhibited a significantly lower growth rate after transfection as shown by cell growth curve and MTT assay and flow cytometry analysis.ConclusionsUsing RNAi technology,siRNA targeting ADAM17 mediated by oligofectamine efficiently knocks down the expression of ADAM17 in human breast cancer MCF-7 cells, results in decrease of cell proliferation activity, arrests cell cycle. RNAi is a new gene silencing technology with high efficiency, ADAM17 can be the candidate genes for gene therapy of breast cancer, which will have potential application prospects in clinics.