Overexpression of KIF2A is Suppressed by miR-206 and Associated with Poor Prognosis in Ovarian Cancer.

BACKGROUND/AIMS: This study aimed to investigate the expression and prognostic value of kinesin family member 2A (KIF2A) and the suppression effects of microRNA-206 (miR-206) on KIF2A in ovarian cancer. METHODS: Ovarian cancer tissues from patients and ovarian cancer cell lines (A2780 and SKOV3) were used in this study. miR-206 mimics and control were transiently transfected into cells. RT-qPCR was performed to detect KIF2A mRNA and miR-206 expression levels, Western blot was performed to detect KIF2A protein levels, Dual-Luciferase Reporter Assay was used to examine the inhibition effects of miR-206 on KIF2A mRNA, immunohistochemical staining was used to examine the expression of KIF2A in tissue sections. CCK-8, transwell and Annexin-V-FITC/Propidium Iodide staining with flow cytometry were used to detect the cell proliferation, migration/invasion, and apoptosis respectively. RESULTS: Our study explored the expression profiles of KIF2A and miR-206 in the patients with ovarian cancer. We found that overexpression of KIF2A was associated with a poor prognosis in ovarian cancer. We also found that KIF2A mRNA contains two target sites for miR-206 binding and confirmed that miR-206 directly suppresses KIF2A; inhibits ovarian cancer cell proliferation, migration, and invasion; and induces apoptosis. CONCLUSION: The results suggest KIF2A could serve a valuable prognostic indicator in ovarian cancer and provide a rationale for treatment of ovarian cancer by targeting KIF2A via miR-206.

Identification of serum microRNA-21 as a biomarker for early detection and prognosis in human epithelial ovarian cancer.

Recent investigations have confirmed up-regulation of serum miR-21 and its diagnostic and prognostic value in several human malignancies. In this study, we examined serum miR-21 levels in epithelial ovarian cancer (EOC) patients, and explored its association with clinicopathological factors and prognosis. The results showed significantly higher serum miR-21 levels in EOC patients than in healthy controls. In addition, increased serum miR-21 expression was correlated with advanced FIGO stage, high tumor grade, and shortened overall survival. These findings indicate that serum miR-21 may serve as a novel diagnostic and prognostic marker, and be used as a therapeutic target for the treatment of EOC.

[Establishment of damaged endometrial stromal cells model in vitro].

OBJECTIVE: To investigate the method of establishing damaged endometrial stromal cells (ESC) model in vitro. METHODS: (1) From June to December 2011 ESC from normal endometrim at proliferation phase (n = 8) and secretory phase (n = 8) were isolated, cultured and identified in vitro. (2) ESC was treated with different concentrations of mifepristone or withdrawal of mifepristone at different time point. The proliferation inhibition percent was measured by cell counting kit-8 (CCK-8). (3) 0 µmol/L(control group)and 60 µmol/L (experimental group) concentration of mifepristone was added into ESC for 48 hours, then withdrew of mifepristone, continued to be cultured for 48 hours. The morphological changes were observed and apoptosis of ESC in different menstrual cycle were detected by flow cytometry. The mRNA and protein level of vascular endothelial growth factor (VEGF), caspase-3, 8, and 9 were determined by one-step quantitative real-time PCR (Q-PCR) and western blot. RESULTS: (1) ESC from 16 specimens of endometrium were all isolated and cultured successfully. (2) The proliferation inhibition rate of ESC was correlated with concentration and duration of mifepristone positively. The proliferation of ESC could be recovered at a range of time after withdrawal of mifepristone. However, when the concentration of mifepristone was 100 µmol/L, the growth of ESC recovered very hardly. (3) The damaged ESC spacing increased, the spindle shape and vacuolization in the cytoplasm were observed in experimental group; the rate of apoptosis of these damaged cells was significantly increased compared with control groups, which were (52 ± 12)% vs. (13 ± 5)% at the proliferative phase and (53 ± 6)% vs. (32 ± 3)% at the secretory phase (all P < 0.05). The relative mRNA level of VEGF was 0.52 ± 0.12 in experimental group and 1.00 ± 0.17 in control group at proliferation phase (P < 0.05). And the relative mRNA level of VEGF was 0.19 ± 0.03 in experimental group and 0.81 ± 0.07 in control group at secretory phase (P < 0.05). The relative level of VEGF protein in the experimental group were both decreased 1.98 and 2.79 folds at the proliferation phase and the secretory phase when compared with those in control group, respectively (P < 0.05). While the relative levels of caspase-3, 8, 9 mRNA were 5.62 ± 0.65, 5.41 ± 0.53, 7.22 ± 0.51 in the experimental group and 1.00 ± 0.44, 1.00 ± 0.21, 1.00 ± 0.32 in control group at the proliferative phase. In the mean time, the relative levels of caspase-3, 8, 9 mRNA were 10.22 ± 0.72, 25.3 ± 1.72, 9.48 ± 1.89 in experimental group and 1.42 ± 0.14, 1.14 ± 0.28, 1.16 ± 0.12 in control group at the secretory phase, respectively (P < 0.05). Compared with the control group, the levels of caspase protein in the experimental group were increased 2.04 and 1.60 folds in caspase-3, 4.23 and 1.49 folds in caspase-8, 2.65 and 3.5 folds in caspase-9 at the proliferative phase and at the secretory phase, respectively (P < 0.05). CONCLUSION: The damaged model of ESC can be established after 48 hours by the withdrawal of 60 µmol/L mifepristone in treatment of ESC for 48 hours.