Construction of eukaryotic vector expressing antisense RNA of ovarian cancer associated antigen— CD147 and clone identification

Objectives To construct eukaryotic antisense RNA expression vector of CD147 and to identify the clones. Methods PCDNAasCD147 was constructed by inserting CD147 cDNA reversely to eukaryotic expression vector PCDNA3.1 .The ovarian cancer cell strain 8910 was transfected by PCDNA as CD147. After selected by G418,positive clones were identified by fluorocytometry assay and indirect immunofluorescence staining. Result It was verified by partial nucleotide sequencing and restriction endonuclease digestion that the constructed eukaryotic antisense RNA expression vector PCDNAasCD147 was correct. Indirect immunofluorescence staining and fluorocytometric assay showed that the expression of CD147 was significantly suppressed in 8910 cells transfected by PCDNAasCD147 as compared with the untransfected 8910 cells. Conclusion The results of this study lay the foundation for further studying the biological functions of CD147 and gene therapy of ovarian cancer.