DNM3OS Enhances the Apoptosis and Senescence of Spermatogonia Associated with Nonobstructive Azoospermia by Providing miR-214-5p and Decreasing E2F2 Expression

Figure 5

Downregulation of miR-214-5p decreased the apoptosis and senescence of spermatogonia induced by ADR treatment. GC-1 spg cells were treated with 0.5 μM ADR for 24 hr and subsequently transfected with the miR-214-5p inhibitor or NC. (a) The levels of DNM3OS and miR-214-5p expression were determined by quantitative real-time PCR. (b) and (c) E2F2 expression at the mRNA and protein levels was determined. (d) Cellular activity was determined by the CCK-8 assay. (e) EDU incorporation assays were performed to assess cell proliferation ability. (f) Apoptotic cells were identified by flow cytometry with Annexin V-PI double staining. (g) Senescence-associated β-galactosidase (SA-β-gal) staining was used to evaluate cell senescence. Data represent the mean value ± SD. ^##, ^### represents NC vs. NC + ADR; , represents NC + ADR vs. miR-214-5p inhibitor + ADR.

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