ABSTRACT
<p><b>OBJECTIVE</b>To study the function of c-myc and K-ras in tumorigenesis of ovarian cancer.</p><p><b>METHODS</b>K-ras and/or c-myc cDNAs were introduced into mouse ovarian surface epithelium cells (MOSE) using recombinant Moloney retroviral vectors. The resulting MOSE cells were studied by cell proliferation assays, the ability to form colonies in soft agarose, matrigel invasion assays and tumorigenicity assays in nude mice.</p><p><b>RESULTS</b>K-ras and c-myc can be easily delivered to the normal MOSE cells by recombinant retroviruses. mRNA and protein of the target genes can be detected by RT-PCR and Western blot. Cell proliferation assays showed that MOSE-Ras cells and MOSE-RM cells (MOSE-Ras/Myc) grew more rapidly than parental cells (MOSE) and MOSE-Myc cells (P <0.01). In addtition, MOSE-RM cells grew more rapidly than MOSE-Ras cells (P <0. 05). Cell colony formation assays showed that while MOSE-Ras and MOSE-RM cells can form colonies in soft-agarose, the MOSE-Myc and MOSE cells did not. Matrigel invasion assays showed that MOSE-Ras and MOSE-RM cells have invasion ability, but not MOSE-Myc ascites and the control MOSE cells. Xenograft experiments showed that MOSE-Ras and MOSE-RM cells were able to form tumors in nude mice following intraperitoneal injection. Tumors were not observed in animals injected with either MOSE-Myc or MOSE cells.</p><p><b>CONCLUSION</b>The recombinant Moloney retroviral system is a highly efficient and convenient method for introducing and expressing foreign genes in murine surface epithelial cell cultures. In this model, expression of K-ras alone is sufficient to generate tumorigenic MOSE, however expression of c-myc in conjunction with K-ras results in cells with a higher index of malignancy. Based on the assays described in this report, expression of c-myc alone can not transform MOSE cultures although it does play a role in cooperation with K-ras.</p>
Subject(s)
Animals , Female , Mice , Blotting, Western , Cell Movement , Cell Proliferation , Cell Transformation, Neoplastic , Cells, Cultured , Epithelial Cells , Cell Biology , Metabolism , Immunohistochemistry , Mice, Nude , Mice, Transgenic , Neoplasms, Experimental , Genetics , Metabolism , Pathology , Oncogene Protein p21(ras) , Genetics , Ovarian Neoplasms , Genetics , Metabolism , Pathology , Ovary , Cell Biology , Proto-Oncogene Proteins c-myc , Genetics , RNA, Messenger , Genetics , Recombinant Proteins , Retroviridae , Genetics , Reverse Transcriptase Polymerase Chain Reaction , TransfectionABSTRACT
Objective To evaluate the relationship between combined muhigene detection and response to chemotherapy and prognosis in epithelial ovarian carcinomas(EOCS).Methods A total of 80 ovarian tissue samples taken from the surgical specimens of patients with EOCS of our hospital in the last two decades who had received chemotherapy "after surgery were paraffin-embedded.The samples were divided into 2 groups,good prognosis group(patients who survived more than 2 years,n=46)and poor prognosis group(patients who survived less than 2 years,n=34).The expression levels of ToPo-Ⅱ,Ki-67,MGMT, PCNA,p27,p53,pl6,P-gp,LRP,GST-?,bcl-2,C-myc,Fas,bax,MSH2,MRP and BCRP were investigated by the combination of tissue arrays and immunohistoehemical streptavidin-biotin peroxidase(SP) method in all samples.Data were analysed with SPSS 12.0 for windows.Results There were statistically significant differences in the positive expression levels of P-gp,BCRP,MGMT,MSH2,p27 and p16(62%, 50% and 50% in poor prognosis group vs 33%,28% and 28% respectively,P