Isocitrate dehydrogenase 2(IDH2)and glutamate dehydrogenase 1(GLUD1)are key enzymes involved in the production ofα-ketoglutarate(α-KG),a metabolite central to the tricarboxylic acid cycle and glutamine metabolism.In...Isocitrate dehydrogenase 2(IDH2)and glutamate dehydrogenase 1(GLUD1)are key enzymes involved in the production ofα-ketoglutarate(α-KG),a metabolite central to the tricarboxylic acid cycle and glutamine metabolism.In this study,we investigated the impact of IDH2 and GLUD1 on early porcine embryonic development following IDH2 and GLUD1 knockdown(KD)via doublestranded RNA(dsRNA)microinjection.Results showed that KD reducedα-KG levels,leading to delayed embryonic development,decreased blastocyst formation,increased apoptosis,reduced blastomere proliferation,and pluripotency.Additionally,IDH2 and GLUD1 KD induced abnormally high levels of trimethylation of lysine 20 of histone H4(H4K20me3)at the 4-cell stage,likely resulting in transcriptional repression of embryonic genome activation(EGA)-related genes.Notably,KD of lysine methyltransferase 5C(KMT5C)and supplementation with exogenousα-KG reduced H4K20me3 expression and partially rescued these defects,suggesting a critical role of IDH2 and GLUD1 in the epigenetic regulation and proper development of porcine embryos.Overall,this study highlights the significance of IDH2 and GLUD1 in maintaining normal embryonic development through their influence onα-KG production and subsequent epigenetic modifications.展开更多
[目的]探讨miR-935靶向GLUD1调控LKB1缺失型肺癌细胞失巢凋亡的潜在机制。[方法]分离条件下培养肺癌细胞构建失巢细胞模型,抽取各个分离时间点的细胞总RNA进行miRNA-seq,过表达或敲低差异miRNA后检测LKB1缺失型肺癌细胞A549和H157的失...[目的]探讨miR-935靶向GLUD1调控LKB1缺失型肺癌细胞失巢凋亡的潜在机制。[方法]分离条件下培养肺癌细胞构建失巢细胞模型,抽取各个分离时间点的细胞总RNA进行miRNA-seq,过表达或敲低差异miRNA后检测LKB1缺失型肺癌细胞A549和H157的失巢凋亡水平。通过miRDB在线分析和遗传学筛选鉴定miRNA的关键底物。根据是否过表达miR-935分为对照组和miR-935过表达组;根据是否敲低GLUD1分为对照组和GLUD1敲低组。[结果]随着分离时间的延长,肺癌细胞中miR-935的表达水平下降(1.47±0.15 vs 0.09±0.01,P<0.05)、GLUD1的表达水平上升(0.87±0.16 vs 1.44±0.21,P<0.05)。过表达miR-935后,肺癌细胞的失巢凋亡水平上升[(15.87±2.23)%vs(49.79±7.63)%,P<0.05]。敲低GLUD1后,肺癌细胞的失巢凋亡水平显著上升[(16.32±3.11)%vs(48.21±5.67)%,P<0.05]。过表达miR-935后,肺癌细胞中GLUD1 mRNA和蛋白的表达水平下降(0.87±0.16 vs 0.10±0.01,P<0.05)。miR-935与GLUD1 mRNA的3′UTR有相互作用。敲低GLUD1后,肺癌细胞内α-酮戊二酸(α-KG)的水平显著下降[(59.32±6.13)μmol/L vs(41.22±3.93)μmol/L,P<0.05]。敲低GLUD1后加入methyl-α-KG,A549和H157细胞的失巢凋亡水平回到了与未敲低GLUD1细胞的相似水平。[结论]miR-935靶向GLUD1 mRNA的3′UTR并减少了GLUD1的表达水平,促进了LKB1缺失型肺癌细胞的失巢凋亡。展开更多
Glutamate dehydrogenase 1(GLUD1)is implicated in oncogenesis.However,little is known about the relationship between GLUD1 and hepatocellular carcinoma(HCC).In the present study,we demonstrated that the expression leve...Glutamate dehydrogenase 1(GLUD1)is implicated in oncogenesis.However,little is known about the relationship between GLUD1 and hepatocellular carcinoma(HCC).In the present study,we demonstrated that the expression levels of GLUD1 significantly decreased in tumors,which was relevant to the poor prognosis of HCC.Functionally,GLUD1 silencing enhanced the growth and migration of HCC cells.Mechanistically,the upregulation of interleukin-32 through AKT activation contributes to GLUD1 silencing-facilitated hepatocarcinogenesis.The interaction between GLUD1 and AKT,as well asα-ketoglutarate regulated by GLUD1,can suppress AKT activation.In addition,LIM and SH3 protein 1(LASP1)interacts with GLUD1 and induces GLUD1 degradation via the ubiquitin–proteasome pathway,which relies on the E3 ubiquitin ligase synoviolin(SYVN1),whose interaction with GLUD1 is enhanced by LASP1.In hepatitis B virus(HBV)-related HCC,the HBV X protein(HBX)can suppress GLUD1 with the participation of LASP1 and SYVN1.Collectively,our data suggest that GLUD1 silencing is significantly associated with HCC development,and LASP1 and SYVN1 mediate the inhibition of GLUD1 in HCC,especially in HBV-related tumors.展开更多
基金supported by the National Research Foundation(NRF)of Korea grant funded by the Korean government(MSIT)(2022R1A2C300769),Republic of Korea。
文摘Isocitrate dehydrogenase 2(IDH2)and glutamate dehydrogenase 1(GLUD1)are key enzymes involved in the production ofα-ketoglutarate(α-KG),a metabolite central to the tricarboxylic acid cycle and glutamine metabolism.In this study,we investigated the impact of IDH2 and GLUD1 on early porcine embryonic development following IDH2 and GLUD1 knockdown(KD)via doublestranded RNA(dsRNA)microinjection.Results showed that KD reducedα-KG levels,leading to delayed embryonic development,decreased blastocyst formation,increased apoptosis,reduced blastomere proliferation,and pluripotency.Additionally,IDH2 and GLUD1 KD induced abnormally high levels of trimethylation of lysine 20 of histone H4(H4K20me3)at the 4-cell stage,likely resulting in transcriptional repression of embryonic genome activation(EGA)-related genes.Notably,KD of lysine methyltransferase 5C(KMT5C)and supplementation with exogenousα-KG reduced H4K20me3 expression and partially rescued these defects,suggesting a critical role of IDH2 and GLUD1 in the epigenetic regulation and proper development of porcine embryos.Overall,this study highlights the significance of IDH2 and GLUD1 in maintaining normal embryonic development through their influence onα-KG production and subsequent epigenetic modifications.
文摘[目的]探讨miR-935靶向GLUD1调控LKB1缺失型肺癌细胞失巢凋亡的潜在机制。[方法]分离条件下培养肺癌细胞构建失巢细胞模型,抽取各个分离时间点的细胞总RNA进行miRNA-seq,过表达或敲低差异miRNA后检测LKB1缺失型肺癌细胞A549和H157的失巢凋亡水平。通过miRDB在线分析和遗传学筛选鉴定miRNA的关键底物。根据是否过表达miR-935分为对照组和miR-935过表达组;根据是否敲低GLUD1分为对照组和GLUD1敲低组。[结果]随着分离时间的延长,肺癌细胞中miR-935的表达水平下降(1.47±0.15 vs 0.09±0.01,P<0.05)、GLUD1的表达水平上升(0.87±0.16 vs 1.44±0.21,P<0.05)。过表达miR-935后,肺癌细胞的失巢凋亡水平上升[(15.87±2.23)%vs(49.79±7.63)%,P<0.05]。敲低GLUD1后,肺癌细胞的失巢凋亡水平显著上升[(16.32±3.11)%vs(48.21±5.67)%,P<0.05]。过表达miR-935后,肺癌细胞中GLUD1 mRNA和蛋白的表达水平下降(0.87±0.16 vs 0.10±0.01,P<0.05)。miR-935与GLUD1 mRNA的3′UTR有相互作用。敲低GLUD1后,肺癌细胞内α-酮戊二酸(α-KG)的水平显著下降[(59.32±6.13)μmol/L vs(41.22±3.93)μmol/L,P<0.05]。敲低GLUD1后加入methyl-α-KG,A549和H157细胞的失巢凋亡水平回到了与未敲低GLUD1细胞的相似水平。[结论]miR-935靶向GLUD1 mRNA的3′UTR并减少了GLUD1的表达水平,促进了LKB1缺失型肺癌细胞的失巢凋亡。
基金supported by Xuzhou Technology Bureau Foundation(KC21065)a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD),the Natural Science Foundation of Jiangsu Province(BK20211347)+2 种基金the National Natural Science Foundation of China(82372245)the Natural Science Foundation of the Jiangsu Higher Education Institutions(21KJA310004)an open Competition Grant of Xuzhou Medical University(JBGS202202).
文摘Glutamate dehydrogenase 1(GLUD1)is implicated in oncogenesis.However,little is known about the relationship between GLUD1 and hepatocellular carcinoma(HCC).In the present study,we demonstrated that the expression levels of GLUD1 significantly decreased in tumors,which was relevant to the poor prognosis of HCC.Functionally,GLUD1 silencing enhanced the growth and migration of HCC cells.Mechanistically,the upregulation of interleukin-32 through AKT activation contributes to GLUD1 silencing-facilitated hepatocarcinogenesis.The interaction between GLUD1 and AKT,as well asα-ketoglutarate regulated by GLUD1,can suppress AKT activation.In addition,LIM and SH3 protein 1(LASP1)interacts with GLUD1 and induces GLUD1 degradation via the ubiquitin–proteasome pathway,which relies on the E3 ubiquitin ligase synoviolin(SYVN1),whose interaction with GLUD1 is enhanced by LASP1.In hepatitis B virus(HBV)-related HCC,the HBV X protein(HBX)can suppress GLUD1 with the participation of LASP1 and SYVN1.Collectively,our data suggest that GLUD1 silencing is significantly associated with HCC development,and LASP1 and SYVN1 mediate the inhibition of GLUD1 in HCC,especially in HBV-related tumors.
