4.5 Article

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

Journal

EXPERIMENTAL AND MOLECULAR PATHOLOGY
Volume 118, Issue -, Pages -

Publisher

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.yexmp.2020.104588

Keywords

LOXL1-AS1; miR-515-5p; NF-kappa B signaling pathway; Choriocarcinoma; Proliferation; Migration

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This study elucidated the molecular mechanism of LOXL1-AS1 in trophoblast cell proliferation and migration, showing that it can target miR-515-5p and NF-kappa B signaling pathways. The results suggest that LOXL1-AS1 may serve as a potential therapeutic target in cancer treatment.
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-LOXL1AS1, 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 Dl, matrix metalloproteinase 2 (MMP2), matrix metalloproteinases 9 (MMP9), phosphorylated p65 (p-p65), profilin of phosphorylated nuclear transcription factor kappa B (p-I kappa B alpha). MTT assay was used to detect cell survival rate, and the luciferase assay was used to detected the targeting relationship of LOXL1-AS1 and miR515-5p. 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 CyclinDl, 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-hd3a in JeG-3 cells showed that miR515-5p could reversed the inhibitory effect of LOXL1-AS1 on NF-x13 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-kappa B signaling pathways.

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