目的探讨具有GTPase结构域、锚蛋白重复序列和PH结构域2的ADP核糖基化因子GAP(with GTPase domain,Anchor protein repeats and ArfGAP of PH domain 2,AGAP2)、离子型谷氨酸受体(Glutamate ionotropic receptor AMPA type subunit 2,GR...目的探讨具有GTPase结构域、锚蛋白重复序列和PH结构域2的ADP核糖基化因子GAP(with GTPase domain,Anchor protein repeats and ArfGAP of PH domain 2,AGAP2)、离子型谷氨酸受体(Glutamate ionotropic receptor AMPA type subunit 2,GRIA2)、代谢型谷氨酸受体结合蛋白(glutamate receptor interacting protein 1,GRIP1)和WNT信号通路中的MCC基因在肿瘤蛋白p53野生型与突变型结直肠癌中的表达差异,并评估其对细胞生物学行为的影响,尤其是AGAP2在p53突变型结直肠癌中的作用机制。方法收集p53野生型和突变型结直肠癌及癌旁组织样本,采用qRT-PCR、Western blot检测AGAP2、GRIA2、GRIP1、MCC表达;通过生物信息学分析AGAP2与GRIA2、GRIP1和MCC的蛋白相互作用,并利用免疫共沉淀技术进行验证;p53突变型结直肠癌细胞株HT29和SW480中敲除AGAP2后用CCK-8检测细胞增殖、Transwell检测侵袭能力,谷胱甘肽(glutathione,GSH)、谷胱甘肽过氧化物酶(glutathione peroxidase,GPX)活力、自由基、Fe 2+检测铁死亡指标。结果与癌旁组织比较,在p53野生型和突变型结直肠癌组织中AGAP2、GRIA2、GRIP1的mRNA和蛋白表达水平均显著上调,而MCC表达显著下调;生物信息学分析表明,AGAP2与GRIA2、GRIP1及MCC具有相互作用,并通过免疫共沉淀实验得到了验证;p53突变型结直肠癌细胞株HT-29和SW480中敲除AGAP2后,CCK-8检测细胞增殖显著降低、Transwell检测侵袭能力受到抑制,铁死亡指标检测发现:细胞中GSH、GPX活力、GPX4蛋白水平均下调,但自由基水平和Fe 2+浓度均显著上调。结论AGAP2在p53突变型结直肠癌中的高表达可能通过调控GRIA2、GRIP1及MCC促进癌细胞的恶性行为。展开更多
Objective The initiation and progression of lung carcinomas are critically regulated by long non-coding RNAs(lncRNAs).However,the role of lncRNAs in the pathways causing lung cancer remains unknown.Methods Cell morpho...Objective The initiation and progression of lung carcinomas are critically regulated by long non-coding RNAs(lncRNAs).However,the role of lncRNAs in the pathways causing lung cancer remains unknown.Methods Cell morphology was regularly observed using an inverted phase-contrast microscope.Cell viability was assessed using CCK-8 according to the manufacturer’s instructions.Total RNA was retrotranscribed from each specimen using the RNAiso Plus Kit.The RT-PCR data were calculated using the Ct approach for comparison.Flow cytometric analyses were prepared by Click-iT™Plus TUNEL Assay for In Situ apoptosis detection,with Alexa Fluor^(TM)594 dye,as instructed.RNA immunoprecipitation assays were used to determine RNA concentration.Results Activated natural killer cells repeat and PH domain-containing protein 2 antisense RNA 1(AGAP2-AS1)levels in cancerous tissues were significantly correlated with cancerous tumor node metastasis(TNM)stage,with cancerous AGAP2-AS1 levels being higher in cancerous tissues than healthy tissues.Patients withelevated AGAP2-AS1 levels had considerably worse outcomes than those with reduced AGAP2-AS1 levels,regardless of the progression-free or overall survival.Functionally,AGAP2-AS1 downregulation represseslung cancer cell growth.AGAP2-AS1 elimination induces erastin-mediated ferroptosis in lung cancer cells.However,the ferritin inhibitor FERSINT-1 negated this result,whereas ERASTIN induced lung cancer cellmortality.After AGAP2-AS1 silencing,erastin-treated lung cancer cells showed a remarkable decrease inGSH levels.These results indicated that AGAP2-AS1 enhanced the stabilization of SLC7A11 mRNA via Recombinant Insulin Like Growth Factor Binding Protein 2(IGF BP2).Patients with elevated AGAP2-AS1 had considerably worse outcomes.Down-regulating AGAP2-AS1 was able to repress lung cancer cell growth and induce greater Erastin-mediated ferroptosis.Lungcancer cells treated with Erastin exhibited a remarkable decrease inglutathione(GSH)levels.The mechanical findingsindicated that AGAP2-AS1 enhanced the stabilization of SLC7A11 mRNA via the IGF2BP2.Conclusion We identified a novel effect of AGAP2-AS1 on TNM staging and the prognosis of patientswith lungcancer by modulating SLC7A11 mRNA stability and ferroptosis.展开更多
Due to the top priority of the command to love God over the command to love one＇s neighbor, these two commands wil/ fall into a profound paradox ＂to violate generally accepted morality for the sake of Christian fait...Due to the top priority of the command to love God over the command to love one＇s neighbor, these two commands wil/ fall into a profound paradox ＂to violate generally accepted morality for the sake of Christian faith. ＂ The orthodox doctrine of ＂justification by faith＂ expresses this paradox in a most definite way, because it regards unbelief in God as the most unacceptable, hateful, and punishable sin against God. Only by giving up this priority can the two love commands truly move beyond the moral paradox and realize their essential and holy unity in the framework of a new ＂ theology of agape＂ or a new doctrine of ＂justification by agape. ＂展开更多
文摘目的探讨具有GTPase结构域、锚蛋白重复序列和PH结构域2的ADP核糖基化因子GAP(with GTPase domain,Anchor protein repeats and ArfGAP of PH domain 2,AGAP2)、离子型谷氨酸受体(Glutamate ionotropic receptor AMPA type subunit 2,GRIA2)、代谢型谷氨酸受体结合蛋白(glutamate receptor interacting protein 1,GRIP1)和WNT信号通路中的MCC基因在肿瘤蛋白p53野生型与突变型结直肠癌中的表达差异,并评估其对细胞生物学行为的影响,尤其是AGAP2在p53突变型结直肠癌中的作用机制。方法收集p53野生型和突变型结直肠癌及癌旁组织样本,采用qRT-PCR、Western blot检测AGAP2、GRIA2、GRIP1、MCC表达;通过生物信息学分析AGAP2与GRIA2、GRIP1和MCC的蛋白相互作用,并利用免疫共沉淀技术进行验证;p53突变型结直肠癌细胞株HT29和SW480中敲除AGAP2后用CCK-8检测细胞增殖、Transwell检测侵袭能力,谷胱甘肽(glutathione,GSH)、谷胱甘肽过氧化物酶(glutathione peroxidase,GPX)活力、自由基、Fe 2+检测铁死亡指标。结果与癌旁组织比较,在p53野生型和突变型结直肠癌组织中AGAP2、GRIA2、GRIP1的mRNA和蛋白表达水平均显著上调,而MCC表达显著下调;生物信息学分析表明,AGAP2与GRIA2、GRIP1及MCC具有相互作用,并通过免疫共沉淀实验得到了验证;p53突变型结直肠癌细胞株HT-29和SW480中敲除AGAP2后,CCK-8检测细胞增殖显著降低、Transwell检测侵袭能力受到抑制,铁死亡指标检测发现:细胞中GSH、GPX活力、GPX4蛋白水平均下调,但自由基水平和Fe 2+浓度均显著上调。结论AGAP2在p53突变型结直肠癌中的高表达可能通过调控GRIA2、GRIP1及MCC促进癌细胞的恶性行为。
基金Supported by the Wuhan Municipal Health Commission Medical Research Project-Youth Project(No.WZ20Q04).
文摘Objective The initiation and progression of lung carcinomas are critically regulated by long non-coding RNAs(lncRNAs).However,the role of lncRNAs in the pathways causing lung cancer remains unknown.Methods Cell morphology was regularly observed using an inverted phase-contrast microscope.Cell viability was assessed using CCK-8 according to the manufacturer’s instructions.Total RNA was retrotranscribed from each specimen using the RNAiso Plus Kit.The RT-PCR data were calculated using the Ct approach for comparison.Flow cytometric analyses were prepared by Click-iT™Plus TUNEL Assay for In Situ apoptosis detection,with Alexa Fluor^(TM)594 dye,as instructed.RNA immunoprecipitation assays were used to determine RNA concentration.Results Activated natural killer cells repeat and PH domain-containing protein 2 antisense RNA 1(AGAP2-AS1)levels in cancerous tissues were significantly correlated with cancerous tumor node metastasis(TNM)stage,with cancerous AGAP2-AS1 levels being higher in cancerous tissues than healthy tissues.Patients withelevated AGAP2-AS1 levels had considerably worse outcomes than those with reduced AGAP2-AS1 levels,regardless of the progression-free or overall survival.Functionally,AGAP2-AS1 downregulation represseslung cancer cell growth.AGAP2-AS1 elimination induces erastin-mediated ferroptosis in lung cancer cells.However,the ferritin inhibitor FERSINT-1 negated this result,whereas ERASTIN induced lung cancer cellmortality.After AGAP2-AS1 silencing,erastin-treated lung cancer cells showed a remarkable decrease inGSH levels.These results indicated that AGAP2-AS1 enhanced the stabilization of SLC7A11 mRNA via Recombinant Insulin Like Growth Factor Binding Protein 2(IGF BP2).Patients with elevated AGAP2-AS1 had considerably worse outcomes.Down-regulating AGAP2-AS1 was able to repress lung cancer cell growth and induce greater Erastin-mediated ferroptosis.Lungcancer cells treated with Erastin exhibited a remarkable decrease inglutathione(GSH)levels.The mechanical findingsindicated that AGAP2-AS1 enhanced the stabilization of SLC7A11 mRNA via the IGF2BP2.Conclusion We identified a novel effect of AGAP2-AS1 on TNM staging and the prognosis of patientswith lungcancer by modulating SLC7A11 mRNA stability and ferroptosis.
文摘Due to the top priority of the command to love God over the command to love one＇s neighbor, these two commands wil/ fall into a profound paradox ＂to violate generally accepted morality for the sake of Christian faith. ＂ The orthodox doctrine of ＂justification by faith＂ expresses this paradox in a most definite way, because it regards unbelief in God as the most unacceptable, hateful, and punishable sin against God. Only by giving up this priority can the two love commands truly move beyond the moral paradox and realize their essential and holy unity in the framework of a new ＂ theology of agape＂ or a new doctrine of ＂justification by agape. ＂
