The Investigation Of Effect Of LON On Cell Characterization By Down Regulation With RNAi In, MCF-7, Human Breast Cancer Cells

The LON/PIMl proteases, localizing in the mitochondria and performing mitochondrial homeostasis and cellular respiration, is essential for both regulatory and housekeeping functions, e.g.: engaged in the process of cell proliferation and differentiation. Under the abnormal conditions such as hypoxia, ER stress, high temperature or brain ischemia, the LON/PIM1 proteases also can regulate the apoptosis of the cells to maintain the normal function of the organism. In this study, we try to down regulate the LON/PIMl proteases in different tumor cells by RNAi technique and check the cell character alteration further. Based on this project, we try to contribute anti-tumor therapy by identifying tumor target molecule.RNA interference (RNAi), which introduce double-stranded RNA (dsRNA) into the cells and lead to the sequence-specific destruction of endogenous RNAs that match the dsRNA, has become a powerful and widely used tool for the analysis of gene function. Whereas, in vivo, longer dsRNA molecules are processed by dicer into 19- to 23nt RNAs, known as short-interfering RNAs (siRNAs), and block the its matched gene expression.For cell model establishing, eight cell lines were quantitated by RT-PCR for LON gene expression and MCF-7, a human breast adenocarcinoma cells, reveal higher expression of LON gene among these tumor cell lines. The short hairpin siRNA sequence of human LON gene was designed with the biological software siRNA Target Finder (Ambion Company). The recombinated plasmid was contruted by ligating these synthesized LON siRNA gene sequence with pSilencer U6 2.1 vector and positive clone,...