目的探究敲减Rho鸟苷三磷酸酶激活蛋白30(Rho GTPase-activating protein 30,ARHGAP30)后,宫颈癌Siha细胞增殖及凋亡的变化。方法设计特异性shARHGAP30引物并连接pLKO.1载体,转化到大肠杆菌感受态细胞中,再与慢病毒辅助质粒转入HEK-293...目的探究敲减Rho鸟苷三磷酸酶激活蛋白30(Rho GTPase-activating protein 30,ARHGAP30)后,宫颈癌Siha细胞增殖及凋亡的变化。方法设计特异性shARHGAP30引物并连接pLKO.1载体,转化到大肠杆菌感受态细胞中,再与慢病毒辅助质粒转入HEK-293T细胞,收集细胞上清获得的病毒过滤后感染Siha细胞,RT-qPCR和Western blot检测敲减效率,以及转染后Bax及Bcl-2的表达变化;CCK-8法检测敲减后细胞的增殖水平。结果成功构建敲减ARHGAP30基因的慢病毒质粒,并建立Siha稳转细胞,ARHGAP30在Siha细胞中的转录和翻译减少(P<0.01),Bax/Bcl-2明显降低(P<0.01),凋亡减少,细胞增殖水平升高(P<0.01)。结论ARHGAP30参与Siha细胞的增殖与凋亡,调控ARHGAP30基因或将干扰宫颈癌的发生和发展。展开更多
Ferroptosis is a pattern of iron-mediated regulatory cell death characterized by oxidative damage.The molecular regulatory mechanisms are related to iron metabolism,lipid peroxidation,and glutathione metabolism.Additi...Ferroptosis is a pattern of iron-mediated regulatory cell death characterized by oxidative damage.The molecular regulatory mechanisms are related to iron metabolism,lipid peroxidation,and glutathione metabolism.Additionally,some immunological signaling pathways,such as the cyclic GMP-AMP synthase-stimulator of the interferon gene axis,the Janus kinase-signal transducer and activator of transcription 1 axis,and the transforming growth factor beta 1-Smad3 axis,may also participate in the regulation of ferroptosis.Studies have shown that ferroptosis is significantly associated with many diseases such as cancer,neurodegenerative diseases,inflammatory diseases,and autoimmune diseases.Considering the pivotal role of ferroptosis-regulating signaling in the pathogenesis of diverse diseases,the development of ferroptosis inducers or inhibitors may have significant clinical potential for the treatment of aforementioned conditions.展开更多
Ferroptosis,an iron-dependent cell death:Ferroptosis is a type of regulated necrosis,characterized by redox-active iron accumulation and increased free radical production derived by Fenton chemistry,that triggers oxid...Ferroptosis,an iron-dependent cell death:Ferroptosis is a type of regulated necrosis,characterized by redox-active iron accumulation and increased free radical production derived by Fenton chemistry,that triggers oxidation of polyunsaturated fatty acids in phospholipids,loss of cellular membranes integrity,and leakage of intracellular contents.展开更多
文摘目的探究敲减Rho鸟苷三磷酸酶激活蛋白30(Rho GTPase-activating protein 30,ARHGAP30)后,宫颈癌Siha细胞增殖及凋亡的变化。方法设计特异性shARHGAP30引物并连接pLKO.1载体,转化到大肠杆菌感受态细胞中,再与慢病毒辅助质粒转入HEK-293T细胞,收集细胞上清获得的病毒过滤后感染Siha细胞,RT-qPCR和Western blot检测敲减效率,以及转染后Bax及Bcl-2的表达变化;CCK-8法检测敲减后细胞的增殖水平。结果成功构建敲减ARHGAP30基因的慢病毒质粒,并建立Siha稳转细胞,ARHGAP30在Siha细胞中的转录和翻译减少(P<0.01),Bax/Bcl-2明显降低(P<0.01),凋亡减少,细胞增殖水平升高(P<0.01)。结论ARHGAP30参与Siha细胞的增殖与凋亡,调控ARHGAP30基因或将干扰宫颈癌的发生和发展。
基金supported by the National Key Research and Development Program of China(Grant No.2022YFA1303900 to S.Y.)the National Natural Science Foundation of China(Grant Nos.32270921 and 82070567 to S.Y.,and 82204354 to Y.H.)+4 种基金the Open Project of State Key Laboratory of Reproductive Medicine of Nanjing Medical University(Grant No.SKLRM-2021B3 to S.Y.)'the Talent Cultivation Project of"Organized Scientific Research"of Nanjing Medical University(Grant No.NJMURC20220014 to S.Y.)the Natural Science Foundation of Jiangsu Province(Grant No.BK20221352 to B.W.)the Jiangsu Provincial Outstanding Postdoctoral Program(Grant No.2022ZB419 to Y.H.)the Postdoctoral Research Funding Project of Gusu School(Grant No.GSBSHKY202104 to Y.H.)the China Postdoctoral Science Foundation(Grant No.2023T160329 to Y.H.).
文摘Ferroptosis is a pattern of iron-mediated regulatory cell death characterized by oxidative damage.The molecular regulatory mechanisms are related to iron metabolism,lipid peroxidation,and glutathione metabolism.Additionally,some immunological signaling pathways,such as the cyclic GMP-AMP synthase-stimulator of the interferon gene axis,the Janus kinase-signal transducer and activator of transcription 1 axis,and the transforming growth factor beta 1-Smad3 axis,may also participate in the regulation of ferroptosis.Studies have shown that ferroptosis is significantly associated with many diseases such as cancer,neurodegenerative diseases,inflammatory diseases,and autoimmune diseases.Considering the pivotal role of ferroptosis-regulating signaling in the pathogenesis of diverse diseases,the development of ferroptosis inducers or inhibitors may have significant clinical potential for the treatment of aforementioned conditions.
基金supported by FONDECYT Initiation in Research,grant number 11201141,awarded to PJU.
文摘Ferroptosis,an iron-dependent cell death:Ferroptosis is a type of regulated necrosis,characterized by redox-active iron accumulation and increased free radical production derived by Fenton chemistry,that triggers oxidation of polyunsaturated fatty acids in phospholipids,loss of cellular membranes integrity,and leakage of intracellular contents.
