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.
Long noncoding RNA (LncRNA) zinc finger protein multitype 2 antisense RNA 1 (ZFPM2-AS1) is highly expressed in a variety of tumors and is involved in promoting the malignant biological behaviors of cancer cells. However, the mechanism of ZFPM2-AS1 in the progression of hepatocellular carcinoma (HCC) remains to be explored. The ZFPM2-AS1 expression in HCC was measured by quantitative real-time PCR (qRT-PCR); cell counting kit-8, 5-bromo-2′-deoxyuridine (BrdU), and transwell assays were used to confirm the biological functions of ZFPM2-AS1 in regulating the malignant biological behaviors of HCC cells; the luciferase reporter gene assay was employed to detect whether ZFPM2-AS1 could bind to microRNA (miR)-576-3p; the regulatory relationship between ZFPM2-AS1 and miR-576-3p was probed by qRT-PCR; the effects of ZFPM2-AS1 and miR-576-3p on the expression of hypoxia-inducible factor 1α (HIF-1α) were detected by qRT-PCR and Western blot. The expression of ZFPM2-AS1 in HCC tissues, compared with that in normal liver tissues, was significantly upregulated. Knockdown of ZFPM2-AS1 markedly inhibited HCC cell proliferation, migration, and invasion while the overexpression of ZFPM2-AS1 worked oppositely. miR-576-3p could reverse the effects of ZFPM2-AS1 on the biological behaviors of HCC cells. Besides, ZFPM2-AS1 could bind to miR-576-3p and positively regulate the expression of HIF-1α, a target gene of miR-576-3p, by adsorbing miR-576-3p. ZFPM2-AS1 is abnormally highly expressed in HCC and facilitates proliferation, migration, and invasion of HCC cells by adsorbing miR-576-3p and upregulating HIF-1α expression.
Background Stereology is an accurate method to obtain 3D quantitative information of microstructure based on the comprehensive observation of two-dimensional sections. Three-dimensional bioprinting of bioink to rebuild a bioengineered ovary has been proved to be a promising technique in preserving fertility. Fully understand the structure of the ovary, the distribution of the main cells and the numerical density of cells using stereology method is of great significance to the computer 3D reconstruction technology.Results The Cavalieri method gave the estimations that the ovary was (2.49 ± 0.32)mm3, the volume fraction of cortex, medulla, follicles and ovarian granulosa cells ( OGCs ) were 86.80%±2.82%, 13.20%±2.82%, 5.60%±0.25% and 81.19%±2.57%, respectively. From exact counts on serial sections, OGCs, primordial, preantral and antral follicles numerical density were estimated at ( 2.11 ± 0.28 ) × 106/mm3, 719.57 ± 18.04/mm3, 71.84 ± 3.93/mm3 and 17.29 ± 3.54/mm3, respectively. We estimated that there would be (2.11 ± 0.28) × 109 OGCs, (719.57 ± 18.04) × 103 primordial follicles, (71.84 ± 3.93) × 103 preantral follicles, (17.29 ± 3.54) × 103 antral follicles in one milliliter 3D bioink when 3D encapsulated mice ovarian constructs. The total number of the three stages follicles per ovary were 1791.74 ± 5.77, 178.88 ± 1.26, 43.04 ± 1.13, and the respective total number of OGCs and follicles per ovary were (5.26 ± 0.09) × 106, 2013.66 ± 8.16.Conclusions OGCs and follicles can be accurately measured using either the optical or physical disector. The application of these approaches for the study of 3D bioengineered ovary may contribute to a deeper understanding of the artificial ovary.
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