b7h3调节髓源性抑制细胞凋亡促进小鼠前列腺癌进展的实验分析 word格式.docxVIP

英文摘要 英文摘要 B7-H3 调节髓源性抑制细胞凋亡促进小鼠前列腺癌进展的实验研究 优秀毕业论文 PAGE PAGE 4 精品参考文献资料 B7-H3 Regulating Myeloid-derived Suppressor Cells Apoptosis in Mouse Prostate Cancer Progression Abstract Objective: We regulated murine prostate cancer RM-1 cells B7H3 gene expression using gene transfection techniques. We observed mouse prostate cancer cell line of RM-1 proliferation in the role of B7H3 by in vivo and in vitro experiments, to explore its possible mechanism. Methods: We retrieved mice B7H3 the nucleotide sequence, synthesized murine B7H3 gene fragment, and transferred to the destination vector, using Gateway Technology. B7H3 gene high expression plasmid vector was constructed. Transfected genes were consistent with the purpose of gene detected by gene sequencing. RT-PCR was used to detect mRNA expression before and after the plasmid construction. The above two methods identified that the plasmids were successfully constructed. Prostate cancer in mice RM-1 cells were stable transfection, and then transfected with plasmid vector B7H3 gene RM-1 cells as a control. Application observed by fluorescence microscopy transfected with green fluorescent protein expression to determine transfection success. After transfection the B7H3 gene expression level was used to detect changes in mRNA expression by RT-PCR. The B7H3 ene expression at the protein level changes in the two groups of cells was detected by flow cytometry. Detection further confirmed that the B7H3 gene was stable transfection. In the part of in-vitro experiments ,the RM-1 cells were divided into two groups: the experimental group (B7-H3 expression group) and negative control group (B7-H3 low expression group only transfected with GFP). Cultured in vitro under the same conditions, the murine prostate cancer cell changes were observated by counting the number of the cells. RM-1 cell growth rate of the two groups were detected for comparing B7H3 gene expression with low B7H3 gene expression in mouse prostate cancer cell gr