High expression of JNK3 for epirubicin-induced apoptosis.docVIP

 PAGE \* MERGEFORMAT 13 High expression of JNK3 for epirubicin-induced apoptosis [Abstract] Objective: To construct c  Jun amino-terminal kinase 3 (JNK3) eukaryotic expression vector transfected into HEK293 cells to establish stable expression cell line; with epirubicin as apoptosis-inducing agent research JNK3 and apoptosis . Methods: pDBLeu  JNK3 plasmid as a template PCR amplification of full-length human JNK3 gene, DNA recombinant technology to its orientation inserted into the eukaryotic expression vector pcDNA3.1/myc  His B, by PCR and restriction enzyme digestion, DNA sequencing, , liposome transfection method transfected HEK293 cells by G418 selection culture, the establishment of stable expression cell lines. RT  PCR, Western Blot Detection of carrying Myc tagged JNK3 fusion protein expression, flow testing of the stable expression of epirubicin JNK3 gene in HEK293 cells and promote apoptosis. Results: pcDNA3.1  JNK3 was successfully constructed the eukaryotic expression vector and has stabilized into HEK293 cells to establish a stable cell line expressing the successful expression of target genes; compared with control cells, compared with epirubicin significantly promote the JNK3 gene highly expressed HEK293 cells apoptosis. Conclusion: JNK3 stable expression cell lines established and high expression of JNK3 for epirubicin-induced apoptosis, in order to further study the function JNK3 provides a good experimental basis. [Keywords:] C  Jum N-terminal kinase expression vector was transfected into eukaryotic cells because of the table 0 Introduction c  Jun amino-terminal kinase (c  Jun N  terminal kinases, JNKs) as the mitogen-activated protein kinase (mitogen activated protein kinase, MAPK) super-families. In vertebrates, JNK by jnk1, jnk2, jnk3 three kinds of gene encoding the expression of JNK1, JNK2, JNK3 protein mainly located in the cytoplasm, in order to serine / threonine protein kinase. JNK3 and JNK1, JNK2 difference is