Translation regulatory long non-coding RNA 1 (TRERNA1) sponges microRNA-23a to suppress granulosa cell apoptosis in premature ovarian failure

Figures & data

Table 1. Participants’ clinical data

Variables	Control (n = 50)	POF (n = 50)
Age (years)	29.66 ± 3.99	29.34 ± 3.89
Basal FSH (IU/L)	6.89 (5.89, 8.13)	15.48 (12.07, 22.37)**
BMI (kg/m2)	23.11 (18.72, 24.34)	22.45 (19.01, 25.99)
Basal LH (IU/L)	5.62 (3.67, 7.99)	5.63 (4.12, 8.99)
Basal E2 (pg/mL)	28.99 (23.23, 41.69)	30.12 (12.11, 42.98)
AMH (ng/mL)	3.21 (2.11, 5.45)	0.48 (0.24, 0.91)**

**p < 0.01.

Figure 1. The expression of TRERNA1 and miR-23a in POF. GC tissue samples donated by both POF patients (n = 50) and controls (n = 50) were analyzed through RNA isolation, followed by RTs and qPCRs to determine the expression of TRERNA1 (A) and miR-23a (B). Each dot represents an average value of three qPCR replicates. Correlations between TRERNA1 and miR-23a across POF samples (C) and control samples (D) were analyzed with Pearson’s correlation coefficient. **p < 0.01.

Figure 2. Detection of TRERNA1 in the subcellular location of granulosa cells and analysis of its interaction with miR-23a. Subcellular fractionation assay was performed to analyze the subcellular location of TRERNA1 in both nuclear and cytoplasm fractions of KGN cells. Two fractions were subjected to RNA isolation, followed by RT-PCR to detect TRERNA1. PCR products were subjected to 1% gel electrophoresis. Images were taken after ethidium bromide staining (A). IntaRNA 2.0 was applied to predict the potential interaction of TRERNA1 with miR-23a (B). RNA-RNA pulldown assay was performed to confirm the interaction between them (C). Data presented were values of mean ±SD of three biological replicates. **p < 0.01.

Figure 3. The role of TRERNA1 and miR-23a in regulating the expression of each other. KGN cells were transfected with TRERNA1 expression vector or miR-23a mimic, and their overexpression was confirmed every 24 h until 96 h. It was observed that TRERNA1 and miR-23a were significantly overexpressed during this time period (A). RT-qPCRs were performed to analyze the role of TRERNA1 in the expression of miR-23a (B) and the role of miR-23a in the expression of TRERNA1 (C). Data presented were values of mean ±SD of three biological replicates. *p < 0.05.

Figure 4. Analysis of the role of TRERNA1 and miR-23a in KGN cell apoptosis. KGN cell apoptosis after the overexpression of TRERNA1 and/or miR-23a was analyzed with cell apoptosis assay.Data presented was values of mean ±SD of three biological replicates. *p < 0.05.