RESUMEN
In this study,real-time PCR and immunohistochemistry were used to detect coxsakie and adenovirus receptor (CAR) expression.Both localization and quantity were evaluated in the uteri obtained at days post coitus (dpc) 2.5,4.5,6.5,8.5.Outcome of PCR was assessed by 2-△△Ct method.Image Pro-Plus 6.0 software was used for quantifying mean density of CAR expression in immunohistochemical sections.We found relatively weak CAR expression in the mouse uteri during implantation window.PCR and immunohistochemistry revealed highest CAR expression was detected on dpc 2.5 followed by down-regulation of CAR at dpc 4.5 and 6.5 (with significant difference).At dpc 8.5,CAR expression was increased slightly again.It is concluded that during implantation,the expression of CAR mRNA and protein is declined,resulting in the impairment of tight junction between cavity epithelium cells.After implantation window closure,CAR appears again to maintain epithelium stability.CAR might play an important role during embryo implantation procedure.
RESUMEN
Full-length coxsackie adenovirus receptor (CAR) eukaryotic expression plasmid was transfected into an ovarian cell line, SKOV3, and its effect on the change of malignant metastasis phenotype was explored. CAR mRNA and protein expression levels among 4 ovarian cancer cell lines (A2780, SKOV3, SW626, CAOV3) and the positive control 293 (a transformed human embryo kidney cell line) was detected by using semi-quantitative RT-RCR and Western blot and compared. CAR-negative SKOV3 was transfected with the eukaryotic expression plasmid containing a full-length CAR cDNA and mock-vector respectively. The positive clones were screened by G418.The biological behavior changes of positive transfected cells were gauged by colony formation in soft agar assay and cell adhesion assay. Among the cell lines, there were obviously different CAR expression levels. CAR could not be detectedin SKOV3. In transfected cell group, CAR expression was enhanced obviously as compared with non-transfected or mock-transfected groups. Cell adhesion in the transfected group was promoted. The number of colony formation was reduced significantly in transfected groups (25.32±8.91) as compared with that in non-transfected group (88.75±13. 98) and mock-transfected group (82. 53 ±19.37). Among the 4 ovarian cancer cell lines,CAR expression level was variable. Exogenous CAR expression had a potential role in inhibiting the malignant metastasis phenotype of ovary cancer cells.