Screening Of HO-1Gene Silencing Stable Transfection Cell Lines Of Gastric Cancer Cell Line SGC7901

Heme oxygenase(HO)is widely present in various tissues of a degradation of hememetabolism rate-limiting enzymes，CO is mainly produced by the metabolism of heme in thebody。It is now thought that oxygen hemoglobin enzyme have three forms：The first type isoxygen stress inducible (HO-1)，the second type is constitutive (HO-2)and the third type is notyet clear，HO-3。As one of heme metabolism rate-limiting enzymes，ho-1will producehemoglobin decomposition of carbon monoxide，Fe2+and biliverdin。Studies have shown that HO-1is not only to play a role in the body’s physiological state，but mainly to play a role in other non-normal state or stress state of the body。A large numberof studies have demonstrated that HO-1is involved in the body of a variety of oxidative stressinjury and inflammation，and play anti-inflammatory，anti-apoptosis and cell protection role。the relationship between ho-1and the tumor takes more and more attention。In recent years，a large number of studies found that HO-1is closely related to tumorproliferation，angiogenesis，apoptosis and other biological behavior。The relationship betweenHO-1and the development of gastric cancer is closely，especially in the role of the process ofperitoneal metastasis，may play a facilitating role，but the mechanism is not yet clear。In order to further in-depth study，we will through the molecular cloning technology，constructed the little interference RNA restructuring expression plasmid carrier，extensivelyproliferate，extraction and purification，and restriction enzyme analysis。Green fluorescence protein(GFP)gene was used as a reporter gene，observed andcompared the key and involved factors in4factors，analysed the efficiency of transfectionand cytotoxicity under different conditions，optimized the condition of transfection。Then we transfected siRNA vector into SGC7901cells，screened HO-1gene silencing stable transfected cell lines using G418for positive screening。Using the methods of RT-PCRand western blot tested and identified the experimental results。We got HO-1gene silencingstable transfection cell lines of gastric cancer successfully。Our results may provide some ideasfor the gene therapy in gastric cancer in future。...