OBJECTIVE To investigate the effect of microR NA-10a on the development of granulosa cells tumor(GCT).METHODS FISH was used to detect the miR-10a expression in tissues from GCT patients.Several functional assays were ...OBJECTIVE To investigate the effect of microR NA-10a on the development of granulosa cells tumor(GCT).METHODS FISH was used to detect the miR-10a expression in tissues from GCT patients.Several functional assays were performed to investigate the effect of miR-10a on proliferation,migration,invasion,spheroid formation and repressed anticancer drug-induced apoptosis of GCT in vitro.CRISPR-Cas9 system mediated miR-10a knockout in cancer GC and two mice GCT models were constructed to show the knockdown effect of miR-10a on cancer GC both in vitro and in vivo.RNA-seq,Western blot,luciferase reporter assay and FISH were used to identify potential direct functional targets and related pathways of miR-10a in cancer GC.RESULTS Strong miR-10a signal was detected in tissues from malignant GCT patients.And amplification of miR-10a negatively correlated with overall survival rate of ovarian cancer patients.In addition,ectopic expression of miR-10a significantly promoted cell proliferation,migration,invasion,spheroid formation and repressed anticancer drug-induced apoptosis in vitro.CRISPR-Cas9 system mediated miR-10a knockout in cancer GC showed opposite phenotype compared to miR-10a overexpressed cancer GC.By using xenograft and orthotropic models,the oncogenic role of miR-10a was further confirmed in vivo.RNA-seq,Western blot,luciferase reporter assay and FISH were used to identified PTEN/TET2 as direct functional targets of miR-10a in cancer GC;Akt and Wnt were found as two associated signaling pathways of miR-10a in cancer GC.CONCLUSION Taken together,our results demonstrate that the miR-10a is positively involved indevelopment of GCT.展开更多
基金supported by Natural Science Foundation of Anhui Province for Young Scholars(1708085QH200)
文摘OBJECTIVE To investigate the effect of microR NA-10a on the development of granulosa cells tumor(GCT).METHODS FISH was used to detect the miR-10a expression in tissues from GCT patients.Several functional assays were performed to investigate the effect of miR-10a on proliferation,migration,invasion,spheroid formation and repressed anticancer drug-induced apoptosis of GCT in vitro.CRISPR-Cas9 system mediated miR-10a knockout in cancer GC and two mice GCT models were constructed to show the knockdown effect of miR-10a on cancer GC both in vitro and in vivo.RNA-seq,Western blot,luciferase reporter assay and FISH were used to identify potential direct functional targets and related pathways of miR-10a in cancer GC.RESULTS Strong miR-10a signal was detected in tissues from malignant GCT patients.And amplification of miR-10a negatively correlated with overall survival rate of ovarian cancer patients.In addition,ectopic expression of miR-10a significantly promoted cell proliferation,migration,invasion,spheroid formation and repressed anticancer drug-induced apoptosis in vitro.CRISPR-Cas9 system mediated miR-10a knockout in cancer GC showed opposite phenotype compared to miR-10a overexpressed cancer GC.By using xenograft and orthotropic models,the oncogenic role of miR-10a was further confirmed in vivo.RNA-seq,Western blot,luciferase reporter assay and FISH were used to identified PTEN/TET2 as direct functional targets of miR-10a in cancer GC;Akt and Wnt were found as two associated signaling pathways of miR-10a in cancer GC.CONCLUSION Taken together,our results demonstrate that the miR-10a is positively involved indevelopment of GCT.
