The Research Of TIMP-1 Gene For The Growth Of Breast Cell

Tissue inhibitor of metalloproteinases-1 (TIMP-1) is a member of the TIMP family, and its main function is inhibition of the activity of various matrix metalloproteinases (MMPs).The association between high levels of protease inhibitor and poor prognosis may be somewhat surprising, However, the recent discovery of other biological functions of TIMP-1 such as growth-stimulating functions, as well as anti-apoptotic and pro-angiogenetic effects, may in part explain this paradox.The purpose of this review is to give an update on the currentstatus of TIMP-1 in breastcancer, emphasizing the prognostic utility of the inhibitor. In addition, the roles of TIMP-1 gene will be outlined.In our experiment, the research of vector were human breast cancer cell (MCF-7) and normal breast epithelial cell(HBL-100). The total RNA was extracted from human breast cancer cell(MCF-7) and the cDNA of the MCF-7 cell was obtained. The TIMP-1 gene length was amplified by the method of normal PCR. The pcDNA3.1-TIMP-1-k and pcDNA3.1-TIMP-1 plasmids were constructed after identity of PCR and restriction, then the plasmids were sequencing by company. Through consequence of TIMP-1 gene was compared to the number sequence of TIMP-1 in the BLAST, then the correct rate was completely 100%. It was demonstrated that we can succeed in constructing the eukaryotic expression plasmid and the interfere RNA plasmid of TIMP-1 gene by company. The human breast cancer cell(MCF-7)and the normal breast epithelial cell (HBL-100) were transfected by the TIMP-1 over-expression plasmid and the TIMP-1 interfere RNA plasmid. After 48-72h, the transcriptional level and protein level were detected by the method of Real time PCR and western blot.It was indicated that the human breast cancer cell(MCF-7)and the normal breast epithelial cell (HBL-100) were transfected by the TIMP-1 over-expression plasmid and the TIMP-1 interfere RNA plasmid. The transcriptional level and protein level was reduced by the TIMP-1 interfere RNA plasmid and.The transcriptional level and protein level was elevated by the TIMP-1 over-expression plasmid. While it is demonstrate that the low level of TIMP-1 can inhibit the growth of cells and over level of TIMP-1 can improve the growth of cells from the growth curve picture. So we can summarize that Promoting human breast cell proliferation is TIMP-1 gene’s function.