ABSTRACT
BACKGROUND: TRIM62 (tripartite motif containing 62) has been found to act as a tumor suppressor of several cancers. However, its precise biological role and related mechanism remain unknown in cervical cancer (CC). METHODS: Quantitative Real-time PCR and western blot were adopted to detect the mRNA and protein expression level of TRIM62 in both human CC cell lines and tissues. Immunohistochemistry was used to measure the TRIM62 expression in 30 normal cervical and 189 CC tissues. Univariate and multivariate Cox regression analyses and Kaplan-Meier survival analyses performed to investigate the association between TRIM62 expression and CC patients' prognosis. The effect of TRIM62 on CC growth and metastasis was studied in vitro and in vivo. Multi-pathway reporter array were utilized to identify the potential signaling manipulated by TRIM62. RESULTS: TRIM62 was frequently down-regulated in both human CC cells and tissues. Low expression of TRIM62 in CC tissues was associated with aggressive clinicopathological features of CC patients. In addition, TRIM62 was also an independent poor prognostic factor for overall and disease-free survival of CC patients after surgery. Moreover, enforced expression of TRIM62 in CC cells significantly inhibited their abilities of proliferation, migration and invasion in vitro. Besides, subcutaneous xenograft tumor model and xenograft mouse metastatic model respectively displayed that TRIM62 impeded the growth and metastasis of CC in vivo. Furthermore, mechanism study exhibited that TRIM62 could suppress epithelial-mesenchymal transition (EMT) by inhibiting c-Jun/Slug signaling. The inhibitory role of TRIM62 in tumor proliferation might be through regulating cell cycle related proteins CyclinD1 and P27 by targeting c-Jun. CONCLUSION: TRIM62 is a potential prognostic biomarker in CC and suppresses metastasis of CC via inhibiting c-Jun/Slug signaling-mediated EMT.
Subject(s)
Proto-Oncogene Proteins c-jun/metabolism , Snail Family Transcription Factors/metabolism , Tripartite Motif Proteins/genetics , Tripartite Motif Proteins/metabolism , Ubiquitin-Protein Ligases/genetics , Ubiquitin-Protein Ligases/metabolism , Uterine Cervical Neoplasms/pathology , Animals , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , Cell Line, Tumor , Cell Movement , Cell Proliferation , Down-Regulation , Epithelial-Mesenchymal Transition , Female , Gene Expression Regulation, Neoplastic , HeLa Cells , Humans , Mice , Neoplasm Metastasis , Neoplasm Transplantation , Prognosis , Signal Transduction , Survival Analysis , Uterine Cervical Neoplasms/genetics , Uterine Cervical Neoplasms/metabolismABSTRACT
OBJECTIVE: To explore the effect of growth differential factor-9 (GDF-9) alone on cell proliferation, cell viability, steroidogenesis, and hormone-stimulated gene expression in cultured mouse theca interstitial cells. DESIGN: Basic research. SETTING: University hospital. ANIMAL(S): Immature 3- to 4-week-old SPF KM mice obtained from the Laboratory Animal Center of Sun Yat-Sen University. INTERVENTION(S): Addition of GDF-9 at different dosages to primary culture of mouse theca interstitial cells. MAIN OUTCOME MEASURE(S): Cell number, cell viability, progesterone and testosterone levels, and hormone-stimulated gene mRNA abundance. RESULT(S): Growth differential factor-9 mildly increased the number of mouse theca interstitial cells and cell viability in a dose-dependent manner and mildly inhibited the production of progesterone in mouse theca interstitial cells. Administration of GDF-9 at the dosages of 200 ng/mL and 400 ng/mL resulted in a significant decrease in the testosterone level compared with the control group by 60.42% and 68.76%, respectively. Growth differential factor-9 significantly suppressed Lhcgr mRNA by 47.36%, Cyp11a1 mRNA by 62.30%, and Cyp17a1 mRNA by 55.39%, but had only a mild effect on Star gene expression. CONCLUSION(S): Growth differential factor-9 can inhibit the production of testosterone in mouse theca interstitial cells and suppress the corresponding gene expression.
