MicroRNAs(miRNAs) have been widely identified in porcine testicular tissues and implicated as crucial regulators of proliferation, apoptosis, and differentiation in porcine spermatogenesis related cells. However, the ...MicroRNAs(miRNAs) have been widely identified in porcine testicular tissues and implicated as crucial regulators of proliferation, apoptosis, and differentiation in porcine spermatogenesis related cells. However, the function roles of most of the miRNAs that have been identified in Sertoli cells are poorly understood. In the present study, six experiments were conducted to study the regulatory role of miR-10b in porcine immature Sertoli cells. In experiment 1, the results showed that the relative mRNA expression level of miR-10b in porcine testicular tissues decreased quadratically(P<0.001) with increasing age, while the relative mRNA expression level of DAZAP1 gene increased(P<0.001). In addition, the mRNA expression of miR-10b was negatively(P<0.01) correlated with DAZAP1 mRNA expression(r=–0.550). In experiment 2, the results from the bioinformatic analysis and a luciferase reporter assay demonstrated that miR-10b directly targeted the DAZAP1 gene in porcine immature Sertoli cells. DAZAP1 mRNA and protein expressions were both regulated(P<0.05) by miR-10b. In experiments 3 to 5, the over-expression of miR-10b or the siRNA-mediated knockdown of the DAZAP1 gene promoted(P<0.05) porcine immature Sertoli cell proliferation, as determined by the Cell Counting Kit-8(CCK-8) assay and the 5-Ethynyl-2′-deoxyuridine(EdU) assay. However, an annexin V-FITC/PI staining assay and the expression of cell survival-related genes indicated that over-expression of miR-10b or knockdown of DAZAP1 had no effect(P>0.05) on porcine immature Sertoli cell apoptosis. In experiment 6, the co-transfection treatment results showed that miR-10b promoted(P<0.05) porcine immature Sertoli cell proliferation by targeting DAZAP1 gene. Overall, these experiments demonstrated that miR-10b promotes porcine immature Sertoli cell proliferation by targeting the DAZAP1 gene.展开更多
Resistance to ferroptosis,a form of regulated cell death caused by disruptions in iron ion and intracellular redox homeostasis,is closely related to tumorigenesis and tumor drug resistance;therefore,targeting ferropto...Resistance to ferroptosis,a form of regulated cell death caused by disruptions in iron ion and intracellular redox homeostasis,is closely related to tumorigenesis and tumor drug resistance;therefore,targeting ferroptosis-related pathways has garnered attention as a potential antitumor therapeutic strategy.However,the molecular mechanisms underlying ferroptosis resistance in tumor cells remain unknown.Zinc-finger estrogen receptor interaction clone 6(ZER6)consists of two isoforms with distinct N-termini,p52-ZER6 and p71-ZER6.ZER6 is upregulated in tumors and promotes tumorigenic potential;however,whether ZER6 is involved in tumor cell ferroptosis resistance remains unknown.Herein,we identified p52-ZER6 as a novel regulator of tumor cell ferroptosis resistance.p52-ZER6 promotes the transcriptional activity of DAZAP1,an RNA-binding protein.DAZAP1,in turn,enhances the stability of SLC7A11 mRNA by binding to its 30-UTR region,thereby increasing SLC7A11 expression and cellular glutathione levels.This subsequently reduces lipid peroxide accumulation and enhances tumor cell ferroptosis resistance,eventually promoting tumorigenic potential.These findings reveal a new function of p52-ZER6 in regulating SLC7A11 mRNA stability via DAZAP1,ultimately leading to ferroptosis resistance and tumorigenic potential.Additionally,we also suggest targeting p52-ZER6 as a potential strategy to promote the efficacy of ferroptosis-based antitumor therapies.展开更多
基金financially supported by the earmarked fund for China Agriculture Research System (CARS-36)the Hunan Provincial Natural Science Foundation of China (2018JJ2176 and 2018JJ3219)
文摘MicroRNAs(miRNAs) have been widely identified in porcine testicular tissues and implicated as crucial regulators of proliferation, apoptosis, and differentiation in porcine spermatogenesis related cells. However, the function roles of most of the miRNAs that have been identified in Sertoli cells are poorly understood. In the present study, six experiments were conducted to study the regulatory role of miR-10b in porcine immature Sertoli cells. In experiment 1, the results showed that the relative mRNA expression level of miR-10b in porcine testicular tissues decreased quadratically(P<0.001) with increasing age, while the relative mRNA expression level of DAZAP1 gene increased(P<0.001). In addition, the mRNA expression of miR-10b was negatively(P<0.01) correlated with DAZAP1 mRNA expression(r=–0.550). In experiment 2, the results from the bioinformatic analysis and a luciferase reporter assay demonstrated that miR-10b directly targeted the DAZAP1 gene in porcine immature Sertoli cells. DAZAP1 mRNA and protein expressions were both regulated(P<0.05) by miR-10b. In experiments 3 to 5, the over-expression of miR-10b or the siRNA-mediated knockdown of the DAZAP1 gene promoted(P<0.05) porcine immature Sertoli cell proliferation, as determined by the Cell Counting Kit-8(CCK-8) assay and the 5-Ethynyl-2′-deoxyuridine(EdU) assay. However, an annexin V-FITC/PI staining assay and the expression of cell survival-related genes indicated that over-expression of miR-10b or knockdown of DAZAP1 had no effect(P>0.05) on porcine immature Sertoli cell apoptosis. In experiment 6, the co-transfection treatment results showed that miR-10b promoted(P<0.05) porcine immature Sertoli cell proliferation by targeting DAZAP1 gene. Overall, these experiments demonstrated that miR-10b promotes porcine immature Sertoli cell proliferation by targeting the DAZAP1 gene.
基金supported by grants from the National Natural Science Foundation of China(82372655,82173029,32270778,and 32070715)the Natural Science Foundation of Chongqing(CSTB2022NSCQ-MSX0611 and CSTB2022NSCQ-MSX0612,China).
文摘Resistance to ferroptosis,a form of regulated cell death caused by disruptions in iron ion and intracellular redox homeostasis,is closely related to tumorigenesis and tumor drug resistance;therefore,targeting ferroptosis-related pathways has garnered attention as a potential antitumor therapeutic strategy.However,the molecular mechanisms underlying ferroptosis resistance in tumor cells remain unknown.Zinc-finger estrogen receptor interaction clone 6(ZER6)consists of two isoforms with distinct N-termini,p52-ZER6 and p71-ZER6.ZER6 is upregulated in tumors and promotes tumorigenic potential;however,whether ZER6 is involved in tumor cell ferroptosis resistance remains unknown.Herein,we identified p52-ZER6 as a novel regulator of tumor cell ferroptosis resistance.p52-ZER6 promotes the transcriptional activity of DAZAP1,an RNA-binding protein.DAZAP1,in turn,enhances the stability of SLC7A11 mRNA by binding to its 30-UTR region,thereby increasing SLC7A11 expression and cellular glutathione levels.This subsequently reduces lipid peroxide accumulation and enhances tumor cell ferroptosis resistance,eventually promoting tumorigenic potential.These findings reveal a new function of p52-ZER6 in regulating SLC7A11 mRNA stability via DAZAP1,ultimately leading to ferroptosis resistance and tumorigenic potential.Additionally,we also suggest targeting p52-ZER6 as a potential strategy to promote the efficacy of ferroptosis-based antitumor therapies.
