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. 2021 Nov 5;14(1):151.
doi: 10.1186/s13048-021-00905-x.

LncRNA DLEU1 is overexpressed in premature ovarian failure and sponges miR-146b-5p to increase granulosa cell apoptosis

Caihong Zheng  1 , Shiwei Liu  1 , Zhihong Qin  2 , Xiaoqian Zhang  3 , Yubao Song  4
Affiliations

LncRNA DLEU1 is overexpressed in premature ovarian failure and sponges miR-146b-5p to increase granulosa cell apoptosis

Caihong Zheng et al. J Ovarian Res. .

Abstract

Background: miR-146b-5p has been reported to participate in premature ovarian failure (POF) in mice. However, its role in POF patients is unclear. We predicted that miR-146b-5p might interact with lncRNA DLEU1, a crucial player in ovarian cancer. We then explored the interaction between DLEU1 and miR-146b-5p.

Methods: Expression of DLEU1 and miR-146b-5p in POF and control ovary tissues was determined by RT-qPCR. The subcellular location of DLEU1 in human KGN cells was analyzed using subcellular fractionation assays. The direct interaction between DLEU1 and miR-146b-5p was analyzed using RNA pull-down assays. The role of DLEU1 in miR-146a expression was analyzed using overexpression assay. Cell proliferation was analyzed using cell apoptosis assay.

Results: Increased DLEU1 expression and decreased miR-146b-5p expression were observed in POF. DLEU1 directly interacted with MiR-146b-5p and was expressed in both nuclear and cytoplasm samples of KGN cells. In KGN cells, DLEU1 and miR-146b-5p failed to regulate the expression of each other. However, DLEU1 promoted cell apoptosis and reduced the inhibitory effects of miR-146b-5p on cell apoptosis.

Conclusions: DLEU1 is overexpressed in POF and sponges miR-146b-5p to increase KGN cell apoptosis.

Keywords: DLEU1; POF; Premature ovarian failure; miR-146b-5p.

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Conflict of interest statement

All authors declare that we do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted.

Figures

Fig. 1
Fig. 1
Exploration of DLEU1 and miR-146b-5p expression in POF. GC samples from both POF patients (n = 49) and controls (n = 49) were used for total RNA isolation. Total RNA samples were subjected to RTs and qPCRs to explore the differential expression of DLEU1 (A) and miR-146b-5p (B) in POF. **, p < 0.01
Fig. 2
Fig. 2
Exploration of the direct interaction between DLEU1 and miR-146b-5p and subcellular location of DLEU1 in KGN cells. IntaRNA 2.0 (A) was used to predict and RNA pull-down assay (B) was performed to validate the direct interaction between DLEU1 and miR-146b-5p. The cellular fractionation assay was carried out to analyze the subcellular location of DLEU1 in KGN cells (C). ***, p < 0.001
Fig. 3
Fig. 3
Exploration of the crosstalk between DLEU1 and miR-146b-5p. Correlations between DLEU1 and miR-146b-5p across both POF (A) and control (B) GC samples were analyzed by Pearson’s correlation coefficient. DLEU1 and miR-146b-5p were overexpressed in KGN cells, and the transfections were confirmed by RT-qPCR every 24 h until 96 h (C). The roles of DLEU1 and miR-146b-5p in the expression of each other in KGN cells were analyzed with RT-qPCRs (D). *, p < 0.05
Fig. 4
Fig. 4
Analysis of the role of DLEU1 and miR-146b-5p in the apoptosis of KGN cells and primary granulosa cells. Cell apoptosis was carried out to study the role of DLEU1 and miR-146b-5p in the apoptosis of KGN cells (A) and primary granulosa cells (B). *, p < 0.05
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