MicroRNAs (miRNAs) have important roles in various types of cellular biological processes. Our study aimed to determine whether miRNAs function in the regulation of ionizing radiation (IR)-induced cell death in auditory cells and to determine how they affect the cellular response to IR. Microarray and qRT-PCR were performed to identify and confirm the differential expression of miRNAs in the cochlea hair cell line HEI-OC1 and in vivo after IR. Upregulation or downregulation of miRNAs using miRNA mimics or inhibitor were detected to characterize the biological effects of the indicated miRNAs. Bioinformatic analyses, luciferase reporter assays and mRNA knockdown were performed to identify a miRNA target gene. We determined that miR-207 was significantly upregulated after IR. MiR-207 enhances IR-induced apoptosis and DNA damage in HEI-OC1 cells. Furthermore, Akt3 was confirmed to be a direct target of miR-207. Downregulation of Akt3 mimics the effects of miR-207. MiR-207 enhances IR-induced apoptosis by directly targeting Akt3 and anti-miR-207 may have a potential role in protecting cochlea hair cells from IR.
ABSTRACT. We explored whether p53 upregulated modulator of apoptosis (PUMA) gene transfection could enhance the sensitivity of epirubicin-induced apoptosis of MCF-7 breast cancer cells. The liposome-mediated recombinant eukaryotic expression vector PU-MA-pCDNA3 and empty vector plasmid were stably transfected into MCF-7 cells. Epirubicin (0.01-100 μM) was applied to MCF-7, MCF-7/PUMA, and MCF-7/pCDNA3 cells for 72 h. The MTT assay was used to calculate the cell survival rate in each group, and the 50% inhibitory concentration (IC50) was calculated. The IC50 values of epirubicin in MCF-7, MCF-7/PUMA, and MCF-7/pCDNA3 cells were 13 ± 1.4, 1.8 ± 0.2, and 10.7 ± 1.3 μM, respectively. The sensitivity of MCF-7/PUMA cells to epirubicin increased 7.2-fold. Epirubicin induced apoptosis in MCF-7 cells dose-dependently, but MCF-7/PUMA cell-induced apoptosis was more significant compared to controls. Low concentrations of epirubicin (0.1 μM) caused low levels of apoptosis of MCF-7/pCDNA3 (1.15 ± 0.26%) and MCF-7 cells (0.9 ± 0.24%), but significantly induced apoptosis of MCF-7/PUMA cells (6.44 ± 1.46%). High epirubicin concentration (1 μM) induced apoptosis in each group, but the epirubicin MCF-7/PUMA apoptosis rate (35.47 ± 9.36%) was significantly higher than that of MCF-7 (12.6 ± 3.73%) and MCF-7/ pCDNA3 (15.2 ± 5.17%) cells (P < 0 01). Flow cytometry and TUNEL assays for apoptosis detection showed similar results. PUMA protein expression in MCF-7/PUMA cells was significantly higher than that in MCF-7 and MCF-7/pCDNA3 cells by Western blot analysis. Therefore, stable transfection of PUMA can significantly enhance epirubicininduced apoptosis sensitivity of MCF-7 breast cancer cells.
Sixty adult male Wistar rats (230-260 g) were randomly divided into 5 equal groups: normal, diabetic, diabetic +800 mg/kg H. hemerocallidea, diabetic+200 mg/kg H. hemerocallidea and non-diabetic+800 mg/kg H. hemerocallidea. Diabetes was induced with a single intraperitoneal injection of 50 mg/kg streptozotocin (STZ) and monitored during the study period. Blood glucose levels showed a significant increase (5.117 ± 0.2412 vs. 30.70 ± 1.2630; p<0.05) 3 days after diabetes was induced using STZ. No mortality was observed during the study period. Blood samples, testes and epididymis were collected on the day of sacrifice. Testicular and epididymal lipid peroxidation (LPO) also showed a significant increase when diabetic control was compared to normal control. Body weights (315.7 ± 9.348 (g) vs. 210.2 ± 6.256 (g); p<0.05), epididymis (1.005 ± 0.0415 (g) vs. 0.7557 ± 0.0279 (g); p<0.05) and testicular (2.995 ± 0.1179 (g) versus 2.140 ± 0.0911 (g); p<0.05) weights, sperm motility and morphology, superoxide dismutase, catalase, total glutathione, total antioxidant capacity, testosterone and estradiol levels (p<0.05) decreased in the diabetic Wistar rats. After 6 week administration of H. hemerocallidea there was a significant improvement in the above parameters in diabetic Wistar rats (p<0.05). In addition, the non-diabetic +800 mg/kg H. hemerocallidea group showed improvement in some sperm motility parameters, epididymal GSHt, testosterone and estradiol levels when compared to the normal control group (p<0.05).These results indicated that H. hemerocallidea supplementation is an effective approach to ameliorate male infertility in diabetic rats.
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