Experimental Study About The Effect Of Trichosanthin On The Epithelial Mesenchymal Transition Of Human Hela Cerval Cancer Cells Through Regulating MiR-1290

Objective To analyze weather trichosanthin(TCS)can effect the epithelial-mesenchymal transition(EMT)of human Hela cervical cancer cells through regulating the expression of miR-1290 and its underlying mechanism.Methods(1)Real-time quantitative PCR technique is used to verify whether the effect of TCS on miR-1290 expression is consistent with the results of the miRNA chip analysis;(2)Transwell technique was used to detect the effect of miR-1290 on the migration and invasion abilities of Hela cells;(3)The effect of miR-1290 on EMT related marker protein Vimentin was detected by Western blot;(4)The target gene of miR-1290 was predicted by data bases miRanda?miRTar Base?miRDB and then target gene RANBP9 was selected as the further research object;(5)The luciferase reporter gene analysis was used to verify the miR-1290 binding sites in RANBP9 m RNA predicted by the software;(6)The effect of miR-1290 on the expression level of RANBP9 was detected by western blot and real-time quantitative PCR.Results(1)Real-time quantitative PCR shown that the down-regulation of miR-1290 expression by TCS was consistent with the chip result;(2)Transwell experiments proved that miR-1290 could promote the migration and invasion of Hela cells;(3)Western blot analysis demonstrated that miR-1290 could promote the expression of Vimentin;(4)The results of luciferase report analysis shown that the 3’UTR of RANBP9 contained the target binding site of miR-1290;(5)Real-time quantitative PCR and western blot assays confirmed that RANBP9 is the downstream target gene of miR-1290.Conclusion Treating Hela cells with Trichosanthin can down-regulate miR-1290 expression and inhibit cervical cancer epithelial mesenchymal transformation through the miR-1290/RANBP9 pathway.