Objective:Circular RNAs(circRNAs)have been shown to involve in pathological processes of ischemic stroke(IS),including autophagy.This study was designed to explore the effect of circR-ZC3HC1 on neuronal autophagy in I...Objective:Circular RNAs(circRNAs)have been shown to involve in pathological processes of ischemic stroke(IS),including autophagy.This study was designed to explore the effect of circR-ZC3HC1 on neuronal autophagy in IS and the related mechanisms.Methods:Expression of circR-ZC3HC1 in blood samples of IS patients and healthy controls was detected.Hippocampal neurons were treated with oxygen and glucose deprivation(OGD)to establish IS in vitro model.The expression of LC3 and p62 and the number of autophagosomes were examined to evaluate the autophagy level of OGD induced neurons using western blotting and transmission electron microscope.Cell apoptosis rate and the expression of cleaved caspase-3,Bax,and Bcl-2 were assessed byflow cytometry and western blotting.The binding relationships among circR-ZC3HC1,miR-384-5p,and SIRT1 were predicted and verified.Results:Low expression of circR-ZC3HC1 was found in blood samples of IS patients and OGD-treated neurons.Overexpressed circR-ZC3HC1 or inhibited miR-384-5p expression promoted autophagy and inhibited apoptosis of OGD-treated neurons,which could be reversed by further 3-MA treatment.Mechanistically,circR-ZC3HC1 targeted miR-384-5p to mediate SIRT1 expression.miR-384-5p overexpression or SIRT1 knockdown in the presence of circR-ZC3HC1 overexpression in OGD-treated neurons lead to reduced autophagy and enhanced apoptosis.Conclusion:Collectively,circR-ZC3HC1 promoted neuronal autophagy to attenuate IS via miR-384-5p/SIRT1 axis.展开更多
目的探讨circKIF4A对卵巢癌SKOV3细胞增殖、凋亡的影响及其对miR-384的调控作用机制。方法采用q RT-PCR法检测卵巢癌组织与癌旁组织中circKIF4A、miR-384的表达量;分别将si-NC、sicircKIF4A、miR-NC、miR-384 mimics、si-circKIF4A与ant...目的探讨circKIF4A对卵巢癌SKOV3细胞增殖、凋亡的影响及其对miR-384的调控作用机制。方法采用q RT-PCR法检测卵巢癌组织与癌旁组织中circKIF4A、miR-384的表达量;分别将si-NC、sicircKIF4A、miR-NC、miR-384 mimics、si-circKIF4A与anti-miR-NC、si-circKIF4A与anti-miR-384转染入SKOV3细胞;采用q RT-PCR法检测SKOV3细胞中circKIF4A、miR-384的表达量;采用MTT实验与流式细胞术分别检测细胞增殖及凋亡能力;双荧光素酶报告实验检测circKIF4A与miR-384的靶向关系。结果与癌旁组织比较,卵巢癌组织中circKIF4A的表达水平升高[(1.00±0.06),(4.28±0.32)](t=62.915, P <0.05),miR-384的表达水平降低[(1.00±0.05),(0.43±0.03)](t=61.047, P <0.05);转染si-circKIF4A后SKOV3细胞OD值降低[(0.75±0.05),(0.41±0.03)](t=17.493, P <0.05),凋亡率升高[(6.36±0.53)%,(23.19±2.21)%](t=22.216,P <0.05);转染miR-384 mimics后SKOV3细胞OD值降低[(0.73±0.05),(0.47±0.04)](t=12.182, P <0.05),凋亡率升高[(7.53±0.41)%,(19.11±1.06)%](t=30.567, P <0.05);双荧光素酶报告实验证实circKIF4A能够吸附miR-384,并可充当miR-384的海绵分子;共转染si-circKIF4A与anti-miR-384后SKOV3细胞OD值升高[(0.40±0.04),(0.65±0.03)](t=15.000, P <0.05),凋亡率降低[(25.20±2.21)%,(10.37±0.86)%](t=18.761, P <0.05)。结论抑制circKIF4A表达可负向调控miR-384的表达从而抑制卵巢癌细胞增殖及诱导细胞凋亡。展开更多
基金Supported by Ningbo Health Technology Project,Nos.2020Y12 and 2022Y12.
文摘Objective:Circular RNAs(circRNAs)have been shown to involve in pathological processes of ischemic stroke(IS),including autophagy.This study was designed to explore the effect of circR-ZC3HC1 on neuronal autophagy in IS and the related mechanisms.Methods:Expression of circR-ZC3HC1 in blood samples of IS patients and healthy controls was detected.Hippocampal neurons were treated with oxygen and glucose deprivation(OGD)to establish IS in vitro model.The expression of LC3 and p62 and the number of autophagosomes were examined to evaluate the autophagy level of OGD induced neurons using western blotting and transmission electron microscope.Cell apoptosis rate and the expression of cleaved caspase-3,Bax,and Bcl-2 were assessed byflow cytometry and western blotting.The binding relationships among circR-ZC3HC1,miR-384-5p,and SIRT1 were predicted and verified.Results:Low expression of circR-ZC3HC1 was found in blood samples of IS patients and OGD-treated neurons.Overexpressed circR-ZC3HC1 or inhibited miR-384-5p expression promoted autophagy and inhibited apoptosis of OGD-treated neurons,which could be reversed by further 3-MA treatment.Mechanistically,circR-ZC3HC1 targeted miR-384-5p to mediate SIRT1 expression.miR-384-5p overexpression or SIRT1 knockdown in the presence of circR-ZC3HC1 overexpression in OGD-treated neurons lead to reduced autophagy and enhanced apoptosis.Conclusion:Collectively,circR-ZC3HC1 promoted neuronal autophagy to attenuate IS via miR-384-5p/SIRT1 axis.
文摘目的探讨circKIF4A对卵巢癌SKOV3细胞增殖、凋亡的影响及其对miR-384的调控作用机制。方法采用q RT-PCR法检测卵巢癌组织与癌旁组织中circKIF4A、miR-384的表达量;分别将si-NC、sicircKIF4A、miR-NC、miR-384 mimics、si-circKIF4A与anti-miR-NC、si-circKIF4A与anti-miR-384转染入SKOV3细胞;采用q RT-PCR法检测SKOV3细胞中circKIF4A、miR-384的表达量;采用MTT实验与流式细胞术分别检测细胞增殖及凋亡能力;双荧光素酶报告实验检测circKIF4A与miR-384的靶向关系。结果与癌旁组织比较,卵巢癌组织中circKIF4A的表达水平升高[(1.00±0.06),(4.28±0.32)](t=62.915, P <0.05),miR-384的表达水平降低[(1.00±0.05),(0.43±0.03)](t=61.047, P <0.05);转染si-circKIF4A后SKOV3细胞OD值降低[(0.75±0.05),(0.41±0.03)](t=17.493, P <0.05),凋亡率升高[(6.36±0.53)%,(23.19±2.21)%](t=22.216,P <0.05);转染miR-384 mimics后SKOV3细胞OD值降低[(0.73±0.05),(0.47±0.04)](t=12.182, P <0.05),凋亡率升高[(7.53±0.41)%,(19.11±1.06)%](t=30.567, P <0.05);双荧光素酶报告实验证实circKIF4A能够吸附miR-384,并可充当miR-384的海绵分子;共转染si-circKIF4A与anti-miR-384后SKOV3细胞OD值升高[(0.40±0.04),(0.65±0.03)](t=15.000, P <0.05),凋亡率降低[(25.20±2.21)%,(10.37±0.86)%](t=18.761, P <0.05)。结论抑制circKIF4A表达可负向调控miR-384的表达从而抑制卵巢癌细胞增殖及诱导细胞凋亡。