ABSTRACT
Objective: To construct eukaryotic expression vector of tumor-associated gene SNC90 and transfect it into human colorectal cancer cell lines. Methods: A 1.5 kb tumor-associate gene SNC90 full length cDNA was inserted into a mammalian expression vector pREP9 to make recombinant vector pREP9-SNC90, which was then introduced into three kinds of colorectal cancer cell lines, SW1116, COL0205 and SW620, by lipofection or electroporation. The cells resistant to G418 drug were selected. Results: Cells transfected with pREP9-SNC90 showed fewer G418-resistant colonies than those transfected with void vector, the inhibitory rates are 72.2%, 74.2% and 59.7%, respectively. Conclusion : SNC90 may play a negative role in regulating growth of colorectal cancer cell.
ABSTRACT
Objective To construct four expression plasmids, pcDNA3.1-GFP/HPV6bE6, pcDNA3.1-GFP/HPV6bE7, pcDNA3.1-GFP/HPV11E6, pcDNA3.1-GFP/HPV11E7 and their transfected murine cell lines. Methods The Four recombinant expression plasmids comprising HPV6bE6,HPV6bE7,HPV11E6 and HPV11E7 linked with GFP, respectively, were constructed and transfected to B16 cells by lipofectamine kit. Positive clones were selected by G418 and observed by fluorescent microscopy and identified by RT-PCR. Results The four constructed recombinant plasmids were authenticated by restriction enzyme digestion and DNA sequencing. Under the fluorescent microscope, the green fluorescence could be observed in cytoplasm and nucleus of four transfected B16 cell lines. The RNA extracted from positively transfected clones resistant to G418 were analyzed by RT-PCR, which demonstrated the presence of four expected fragments. Conclusions The transfected murine cell lines B16 can express HPV6bE6,HPV6bE7,HPV11E6 and HPV11E7 gene. These transfected cell lines can be further transplanted to mice in order to investigate the biological properties and immunological mechanisms of these genes in vivo.
