Targeted-regulating of miR-515-5p by LncRNA LOXL1-AS1 on the proliferation and migration of trophoblast cells

Abstract

Objective: This study aims to elucidate the molecular mechanism of LOXL1-AS1 in proliferation and migration of trophoblast cells.

Methods: We have used specific small interfering RNAs (si-RNA) and microRNA (mi-RNA) to knock down the target gene or m-RNA. In this regard we used following siRNAs: si-NC, si-LOXL1-AS1, pcDNA-NC, pcDNA-LOXL1-AS1, miR-NC, miR-515-5p. These si-RNA and miRNA were transfected into JeG-3 cells. Real-time quantitative PCR (RT-qPCR) was used to detect the expressions of LOXL1-AS1 and miR-515-5p. Western blot was used to detect Cyclin D1, matrix metalloproteinase 2 (MMP2), matrix metalloproteinases 9 (MMP9), phosphorylated p65 (p-p65), profilin of phosphorylated nuclear transcription factor κB (p-IκBα). MTT assay was used to detect cell survival rate, and the luciferase assay was used to detected the targeting relationship of LOXL1-AS1 and miR-515-5p.

Results: Our results showed that human placental trophoblast cells had higher level of LOXL1-AS1 in comparison to human choriocarcinoma cells, however, human placental trophoblast cells had lower level of miR-515-5p. In addition, the expression of CyclinD1, MMP2, MMP9 were significantly decreased in JeG-3 cell lines. We observed lower cell survival rate and lower cell migration number in JeG-3 cell lines. Our results demonstrated that LOXL1-AS1 could target miR-515-5p, and subsequently reverse the inhibitory effect of LOXL1-AS1 on proliferation and migration in JEG-3 cells. Also, lower expressions of p-p65 and p-IкBα in JeG-3 cells showed that miR-515-5p could reversed the inhibitory effect of LOXL1-AS1 on NF-κB signaling pathway.

Conclusions: The low expression of LOXL1-AS1 inhibits the proliferation and migration of human choriocarcinoma cells, which might be related to miR-515-5p and NF-κB signaling pathways.

Keywords: Choriocarcinoma; LOXL1-AS1; Migration; NF-κB signaling pathway; Proliferation; miR-515-5p.