Human KiSS1 and p27Kip1 gene in osteosarcoma MG63 cell lines co-expression.docVIP

 PAGE \* MERGEFORMAT 13 Human KiSS1 and p27Kip1 gene in osteosarcoma MG63 cell lines co-expression [Abstract] Objective To clone the tumor metastasis suppressor gene KiSS 1 and cyclin dependent kinase inhibitor p27Kip1 expression of gene sequences, application of the internal ribosomal insertion site (IRES) sequences were constructed expression vector pIRES KiSS 1 p27Kip1, sequencing, and access to stable cell lines express the target protein. Methods RT PCR from normal human placenta obtained KiSS 1 and p27Kip1 gene open reading frame of the cDNA sequence, cDNA sequences of the two recombinant target gene to the pIRES vector, co-expression vector pIRES KiSS 1 p27Kip1, application of liposome osteosarcoma MG63 cells, and by Western blot, RT PCR and protein expression. Results PCR, restriction enzyme digestion and gene sequencing, the recombinant pIRES vector into the two fragments were open reading frame for the target gene nucleotide sequence, KiSS 1 and p27Kip1 protein in cells transfected with expression. Conclusion The construction of co-expression vector pIRES KiSS 1 p27Kip1, stable expression of KiSS 1 and p27Kip1 protein in cell lines To further observe the gene on human osteosarcoma cells in vitro and in vivo effects of foundation. [Key Words] molecular cloning of genetic recombination osteosarcoma tumor suppressor gene in cell cycle proteins cultured tumor cells KiSS 1 gene is a tumor metastasis suppressor gene, and MMP 9 gene through interaction to increase the adhesion between cells and inhibit tumor metastasis [1 2]. P27Kip1 protein is a cyclin dependent kinase (Cdk) in the negative regulatory factors, reported in osteosarcoma and other malignant tumors, its expression was decreased and associated with a variety of malignant tumors [3]. to study the two genes in the inhibition of proliferation and metastasis of osteosarcoma in the existence of synergies , the author and the p27 gene was cloned KiSS 1, through the eukaryotic expression vecto