| Objectives To investigate the role of miR-101 on the anoikis resistance of epithelial ovarian cancer SKOV3 cells,and the possible mechanisms of this process.Methods 1 SKOV3 cells anoikis pattern was established to using Poly-HEMA solution coated plate;2 miR-101 over-expressing were constructed in SKOV3 cells by cell transfection technique,and the transfection efficiency of SKOV3 cells and the expression level of c-fos m RNA were detected by RT-PCR;3 After miR-101 over-expressing cells were successfully constructed,Transwell invasion assay was used to detect the invasion ability of the cells which were detected;CCK-8 Kit was used to detect the proliferation ability of cells;The wound-healing assay was used to detect the migration ability of cells;The concentration of intracellular Ca2+ and apoptosis rate were detected by Flow Cytometry;Western blot was used to detect the expression level of PKC,p-CREB and cfos protein;Also,The ultrastructure of cells were observed by Transmission Electron Microscope.4 Analysis of variance followed by SNK-q test and repeated measurement data were performed using SPSS 17.0.Results 1 The anoikis pattern was successfully established,and the SKOV3 cells were round,suspended and consolidated into multicelluar spheroids.2 After transfection,Compared with blank control group,mock group and miR-NC group,mi R-101 expression level of SKOV3 cells treated with miR-101 mimicis has been raised significantly in miR-101 plasmid group（P<0.05）,and the other 3 groups were respectively compared,the difference was not statistically significant（P>0.05）;3 After miR-101 over-expressing cells were successful constructed,compared with blank control group,mock group and miR-NC group,the cells invasion ability were decreased in miR-101 plasmid group（P<0.05）;with the increase of time,the proliferation and migration ability in miR-101 plasmid group were significantly lower than the other 3 groups（P<0.05）,and the invasion,proliferation and migration ability of the other 3 groups were not statistically significant（P>0.05）;4 After miR-101 over-expressing cells were successful constructed,compared with blank control group,mock group and miR-NC group,the concentration of Ca2+ was significantly increased in miR-101 plasmid group（P<0.05）,and the other 3 groups were not statistically significant（P>0.05）;5 After miR-101 over-expressing cells were successful constructed,compared with blank control group,mock group and miR-NC group,the expression levels of PKC,p-CREB,c-fos protein and c-fos m RNA were significantly decreased（P<0.05）,and the other 3 groups were not statistically significant（P>0.05）;6 After miR-101 over-expressing cells were successful constructed,the apoptosis rate of miR-101 plasmid group was significantly higher than that of blank control group,mock group and miR-NC group（P<0.05）,and the other 3 groups were not statistically significant（P>0.05）.Conclusions 1 miR-101 could mediate anoikis resistance in epithelial ovarian Cancer cells;2 miR-101 could mediate anoikis resistance in epithelial ovarian cancer cells through the signaling pathway of PKC/CREB/c-fos. |
