miR-486-5p inhibits invasion and migration of HTR8/SVneo trophoblast cells by down-regulating ARHGAP5.

INTRODUCTION: Appropriate implantation of trophoblast cells is necessary for successful pregnancy outcome. This process requires proper migration and invasion of trophoblast cells into the maternal endometrium and the myometrium. Dysregulation of circulating microRNAs in preeclampsia has been reported in several studies. Furthermore, miR-486-5p was reportedly increased within exosomes derived from maternal circulation in preeclamptic pregnancy. However, the roles of elevated miR-486-5p in preeclampsia has not yet been clarified. METHODS: HTR8/SVneo trophoblast cells were transfected with miR-486-5p, and the ARHGAP5 expression was examined by quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blotting. A reporter assay using a luciferase construct containing the ARHGAP5 3'-untranslated region (3'UTR) was performed to determine whether or not ARHGAP5 is a direct target of miR-486-5p. Changes in migration and invasion abilities were examined by a wound healing assay and invasion assay, respectively. RESULTS: The ARHGAP5 expression was significantly decreased in miR-486-5p-transfected cells according to RT-qPCR and Western blotting. A dual luciferase reporter gene assay showed that miR-486-5p acts directly on the 3'UTR of ARHGAP5 mRNA. The migration and invasion abilities were suppressed in miR-486-5p-transfected cells. Downregulation of ARHGAP5 by small interfering RNA transfection inhibited trophoblast cell migration and invasion, resembling that of miR-486-5p transfection. DISCUSSION: The migration and invasion abilities of HTR8/SVneo cells were suppressed by miR-486-5p at least partly through inhibiting the ARHGAP5 expression. These data suggest that miR-486-5p is involved in the pathogenesis of preeclampsia and that miR-486-5p is a viable potential biomarker for predicting the onset risk of preeclampsia.

Outcome of in vitro fertilization cycles with automatic time-lapse instrumentation combined with the early embryo viability assessment score.

AIM: To evaluate whether embryo selection using the early embryo viability assessment (EEVA) score increases the ongoing pregnancy rate of in vitro fertilization and intracytoplasmic sperm injection patients. METHODS: One hundred eighty-one patients whose serum anti mullerian hormone (AMH) level was greater than 0.5 ng/µL were enrolled in the study. All patients received oocyte retrieval repeatedly from June 2017 to January 2019. Transferred embryos were selected using the EEVA score and Veeck's criteria. We investigated the blastocyst rate according to the EEVA score and Veeck's criteria and also evaluated the clinical outcome following embryo transfer of the blastocysts. RESULTS: Blastocyst development rate (48.7%) and high-quality blastocyst (42.4%) of Veeck 1 was statistically higher than others. The blastocyst rate (71.4%) and high-quality blastocyst rate (60.0%) for EEVA 1 was the highest, and a correlation between the EEVA score and the blastocyst rate was also identified in cases younger than 40 years. Blastocyst rate of EEVA 1 + 2 (69.8% 208/298) was statistically higher than that of Veeck 1 + 2 (40.1% 317/791) (p < 0.05) and high-quality blastocyst rate of EEVA 1 + 2 (50.0% 104/208) was also higher than that of Veeck 1 + 2 (36.6% 117/320) (p < 0.05). However, there was a significant correlation between EEVA and the pregnancy rate and pregnancy rate of EEVA 1 + 2 showed no statistical difference compared with Veeck 1 + 2. CONCLUSIONS: Although it remains to be answered whether a computer can substitute Veeck's classification, the EEVA score could be a viable alternative to predict the blastocyst rate and to select those high-potential embryos that improve the pregnancy rate.

The differential expression of miRNAs between ovarian endometrioma and endometriosis-associated ovarian cancer.

BACKGROUND: MicroRNAs (miRNAs) have been implicated to play a vital role in development, differentiation, cell proliferation and apoptosis. However, which miRNAs are actually associated with endometriosis-associated ovarian cancer remains controversial. METHODS: Serum and ascites samples were obtained from all patients. Serum samples from 5 cases of ovarian endometrioma and endometriosis-associated ovarian cancer each were submitted for comprehensive miRNA microarray profiling. We investigated the differential expression of miRNAs between the two groups to confirm the pivotal role of miRNAs. Quantitative reverse transcription-polymerase chain reaction validation of five selected miRNAs [miR-92a-3p, miR-486-5p, miR-4484, miR-6821-5p, and miR-7108-5p] was performed, and miR-486-5p expression analysis was followed by proliferation and wound healing assays, depending on the expression of miR-486-5p. RESULT: miR-486-5p expression in serum and ascites samples from endometriosis-associated ovarian cancer patients was significantly higher than that from ovarian endometrioma patients. Moreover, the miR-486-5p level in serum and ascites samples was significantly correlated with the severity of the endometriosis. The upregulation of miR-486-5p in immortalized ovarian endometrioma cells significantly increased proliferation and migration. In contrast, the downregulation of miR-486-5p in these cells significantly decreased proliferation and migration. CONCLUSION: miR-486-5p might function as an oncogenic miRNA in endometriosis-associated ovarian cancer and could be a noninvasive biomarker to prospect the severity of ovarian endometrioma.