Normal and phosphorylated forms of JNK1 were not affected by TRIM62 (Fig. 5f). These results

Normal and phosphorylated forms of JNK1 were not affected by TRIM62 (Fig. 5f). These results indicated the inhibitory function of TRIM62 on MAPK/JNK signaling was through suppressing the expression of c-Jun. With all these results taken together, it is Tirabrutinib custom synthesis concluded that TRIM62 could suppress EMT by inhibiting c-Jun/ Slug signaling in CC.Rescue of c-Jun abrogates inhibition of TRIM62 on cell migration and invasionHaving demonstrated TRIM62 could suppress EMT by inhibiting c-Jun/Slug signaling, we wanted to further clarify whether the inhibition effect of TRIM62 on cell invasion and migration was through suppressing c-Jun/Liu et al. Journal of Experimental Clinical Cancer Research (2016) 35:Page 14 ofFig. 4 (See legend on next page.)Liu et al. Journal of Experimental Clinical Cancer Research (2016) 35:Page 15 of(See figure on previous page.) Fig. 4 TRIM62 impedes growth and metastasis of cervical cancer in vivo. Effect of TRIM62 on tumor growth and metastasis were investigated in nude mouse xenograft model, which was built by injecting stable CC cells (SiHa-NC, SiHa-TRIM62, HeLa-NC and HeLa-TRIM62). **, P < 0.01; ***, P < 0.001. a A representative picture of tumors originated from nude mice, which had subcutaneously inoculated with stable CC cells as described above for 30 days. b Diagrams showed the calculation of the tumor weight of TRIM62 overexpressed groups (SiHa-TRIM62 and HeLa-TRIM62) compared with that of the corresponding control groups (SiHa-NC and HeLa-NC) (n = 6). c Subcutaneous tumor growth curves of each group were displayed (n = 6). d Typical H E images of pulmonary metastatic foci in control groups and normal pulmonary tissues in TRIM62 overexpressed groups were shown, which indicating the metastatic capacity was decreased on TRIM62 overexpressed groups. Original magnification: ?00, ?00 and ?400. e The diagrams showed the percentages of mice with or without metastatic foci in the lungs (n = 6)Slug signaling-mediated EMT. We transfected c-Jun overexpression lentivirus into TRIM62-overexpressed cells, and the rescue of c-Jun expression in these cells was validated by western blot (Fig. 6a). Firstly, the expression level of -Catenin was significantly decreased after overexpression of c-Jun in TRIM62-overexpressed cells. While the expression level of Vimentin was significantly increased after overexpression of c-Jun in TRIM62-overexpressed cells (Fig. 6b). Then cell function assays were performed. We detected cell morphology with rhodamine-phalloidin fluorescent staining. It demonstrated that epithelial phenotype-like morphology of TRIM62-overexpressed cells changed back to mesenchymal phenotype-like morphology after rescuing the expression of c-Jun (Fig. 6c). Morever, transwell migration and transwell invasion assays were further adopted. Results showed the inhibited migration and invasion by gain of TRIM62 were significantly abrogated after overexpression of c-Jun in TRIM62-overexpressed cells (Fig. 6d). All these results suggested rescue of c-Jun could abrogate inhibition of TRIM62 on cell migration and invasion.TRIM62 regulates the expression of CyclinD1 and P27 via targeting c-Jun in cervical cancersignaling by down-regulating c-Jun. In addition, MAPK/ JNK signaling also has been reported to participate in cell proliferation and tumor growth, and CyclinD1 and P27 could PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/28878015 be regulated by c-Jun [34, 35]. So we hypothesized that these changes caused by TRIM62 were through targeting c-Jun. We detected the expression level of.