2.Effect of hepatocyte growth factor gene transfection on biological features of lymphoma cells.
Dong CEN ; Hang ZHAO ; Rong-rong SHEN ; Shi-xuan HUA ; Jian-xin LU ; Ren-zhi PEI ; Zhi-guang TU
Journal of Zhejiang University. Medical sciences 2010;39(4):378-385
OBJECTIVETo investigate the biological effect of hepatocyte growth factor (HGF) on HGF gene-transfected Raji cells.
METHODSTotal RNA was extracted from human hepatic tissue, HGF gene cDNA was amplified by RT-PCR, and then cloned into vector pVITRO2-mcs to construct recombinant eukaryotic expression vector pVITRO2-mcs-HGF. The recombinant vector was transfected to Raji cells, and the stably transfected cells were selected by homomycin B in serial passages, and confirmed by real-time fluorescent quantitative PCR, ELISA, immunocytohistochemistry. The biological features of transfected Raji cells were evaluated by semisolid culture.
RESULTSRT-PCR results showed that Raji cells were transfected successfully with recombinant eukaryotic expression vector pVITRO2-mcs-HGF. HGF mRNA and protein were expressed successfully in Raji cells. Expression of HGF gene enhanced proliferation, metastasis and invasion of Raji cells.
CONCLUSIONHGF gene has been cloned and recombined to construct recombinant eukaryotic expression vector pVITRO2-mcs-HGF successfully. Transfected HGF may change the biological features of Raji cells.
Cell Line, Tumor ; Cloning, Molecular ; Hepatocyte Growth Factor ; biosynthesis ; genetics ; Humans ; Lymphoma, B-Cell ; genetics ; pathology ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection