Objective:1explore the expression level of FoxM1in clinical gastric cancer specimen2explore the impact on expression of BCL-2by overexpression of FoxM13explore the impact on the gastric cancer cell line proliferation ability by regulation on FoxM1Methods:1abstract the RNA from human blood and perform RT-PCR, using the acquired cDNA library as the template and design primers according to the sequence information on NCBI website, get the sequence of CDS part of FoxM1gene using PCR technology, amplify the CDS sequence with the help of pMD-18T vector, cut the CDS sequence using restricted enzyme and insert it into the pMIG plasmid vector, by penicillin selection,we acquire the FoxM1overexperssion plasmid2transfect the gastric cancer cell line with the acquired FoxM1overexpression plasmid as well as the FoxM1siRNA,harvest the cells and abstract the RNA,performing RT-PCR to get the cDNA library, using the Quantitative-PCR to detect the mRNA expression level of FoxM1and BCL-2, also using western blot to detect the protein expression level of FoxMl and BCL-23transfect the gastric epithelium cancer cell line with the FoxM1overexpression plasmid and FoxM1siRNA and performing the colony formation analysis, revealing the FoxMl impact on the gastric cancer proliferation ability4abstract RNA from the8clinical primary gastric cancer specimen performing RT-PCR to acquire the corresponding cDNA library, using Quantitative-PCR to detect the expression level of FoxM1and BCl-2Results:1using Quantitative-PCR and Western Blot, we validated that the construction of the FoxM1overexpression plasmid succeeded2BCL-2expression level was significantly elevated by the transfection of FoxM1overexpression plasmid in gastric cancer cell line.3gastric cancer cell line gains more proliferation ability after FoxM1is overexpressed.4gastric cancer cell line transfected by FoxM1siRNA has a low BCL-2expression level and lower colony formation ability.58pairs of clinical primary gastric cancer specimen have high FoxM1and BCL2expression levelConclusion:①The transcriptional factor FoxM1has positive correlation with BCL-2②FoxM1could significantly promote the gastric cancer cell proliferation ability by elevating BCl-2expression;③FoxM1and BCL2have higher expression level in clinical primary gastric cancer specimen... |