Effect of miR-7 targeted regulation of CTSK on proliferation and migration of ovarian granulosa cells in Guizhou Capra hircus
[Objective]The interaction between miR-7 and CTSK genes and their effects on the proliferation and apoptosis of ovarian granulosa cells in Capra hircus were explored,so as to provide a basis for the preservation,breeding,development and utilization of good breeds of C.hircus.[Method]The online database was used to screen the binding targets of miR-7 and CTSK gene 3'UTR,and the interaction sites were detected by inserting the CTSK gene 3'UTR into the dual luciferase reporter system.CCK-8 and scratch test were used to detect whether miR-7 targeted CTSK gene to affect the proliferation and migration of ovarian granulosa cells in C.hircus.After overexpression of miR-7,RT-qPCR was used to detect its effect on the expression levels of PCNA,BCL2,BAX,caspase-3 and caspase-9.[Result]The study successfully constructed wild-type and mutant dual-luciferase reporter vectors for the 2 predicted targets in the 3'UTR region of the CTSK gene.The re-sults of dual-luciferase assay showed that the 2 predicted targets in the 3'UTR region of the CTSK gene were regulated by miR-7.The results of CCK-8 and cell scratch showed that the proliferation and migration of the ovarian granulosa cells of the test group transfected with miR-7 were significantly higher than the control group.RT-qPCR results showed that up-regulation of miR-7 could significantly up-regulate the ex-pression of PCNA and BCL2,and inhibit the expression of BAX,caspase-3 and caspase-9.[Conclusion]miR-7 can target two target sites in the 3'UTR region of CTSK and inhibit the expression of CTSK gene.Overexpression of miR-7 can significantly promote the proliferation and migration of ovarian granulosa cells in C.hircus,and promote the expression of proliferation marker genes,anti-apoptotic genes and inhibit the expression of pro-apoptotic genes.The related research results can provide a basis for further study on the molecular mechanism of miR-7 tar-geted regulation of CTSK expression affecting the reproductive performance of C.hircus.
Capra hircusmiRNACTSKOvarian granulosa cellsCell proliferation and migration
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