The Effects On The Biological Characteristics Of Human Prostate Cancer Cells(PC3) After RNA Interfere With The Expression Of Bloom Helicase Gene

DNA helicases exist almost in all cells, they play an important role in the process of cell metabolism and so on. RecQ family helicases, a significant family member of DNA helicases, play a key role in cell metabolism including DNA repair, recombination, transcription, and telomere maintenance. Bloom helicase(BLM) is an important member of RecQ family helicases, it is involved in the process of DNA replication and recombination and so on, it also plays an important role in holding the stability of telomeres and chromosomes. It can interact with a variety of protein factors to affect the function of the factors. The mutation of BLM helicase results in the emergence of BLM syndrome(BS), BS is a rare, recessive autosomal inheritance disorder, whose patients display the liability of many types of cancers. BLM is involved in the cellular response to DNA damage and stalled replication forks in the processes of DNA repair and apoptosis. DNA repair factor such as BLM helicase-specific inhibition in malignant cells may compromise the high proliferation rates of cancerous cells. The role of RecQ helicase suggests that the helicase may be as a molecular target for anti-cancer agents. The research applied comprehensively various methods and techniques of biochemistry,molecular biology, cell biology to construct the RNAi vector of BLM helicase, and studied the effect of the interference vector on cell biological characteristics of human prostate cancer PC3 cells. BLM helicase will probably be a new target of anti-cancer, which will provide new idea and new target of drugs action. The results as follow,1. According to the Bloom helicase gene sequence on NCBI, based on design principle of shRNA to designed and synthesized of two RNAi expression vectors of BLM helicase gene, then the vectors were transformed into E.coli competent cells to clone the vectors. By agarose gel electrophoresis and DNA sequencing confirmed the successful constructionof the recombination vectors. It can be used for the next step experiment.2. Fluorescence quantitative PCR technology conformed that the RNAi expression vectors of BLM helicase gene could effectively inhibit the BLM helicase gene expression from gene level, Western blot proved the RNAi vectors could effectively inhibit the expression of BLM helicase protein from the protein level.3. Through MTT assay, scratch assay and Transwell assay attested to the interference vectors of BLM helicese could inhibit the proliferation,migration and invasion ability of PC3 cells significantly, Hoechst / PI experiment proved that the vectors played a significant role in promoting the apoptosis rate of PC3 cells.