To study the roles and association of murine double minute gene 2(MDM2)and fructose-1,6-biphosphatase(FBP1)in human hepatocellular carcinoma(HCC),growth response of human HCC cells was assessed using proliferation and...To study the roles and association of murine double minute gene 2(MDM2)and fructose-1,6-biphosphatase(FBP1)in human hepatocellular carcinoma(HCC),growth response of human HCC cells was assessed using proliferation and apoptosis assay.Pro-survival AKT signaling associated proteins(p-AKT,survivin and cleaved caspase 3)were assessed using western blotting.The correlation between MDM2 and FBP1 was assessed using coimmunoprecipitation combined with ubiquitination assay.Our data suggested that low expression of FBP1 was correlated with high levels of MDM2 in HCC cell lines(Huh7 and Hep3B).Overexpression of FBP1 resulted in antiproliferation,pro-apoptosis,the up-regulation of cleaved caspase 3 while the downregulation of survivin and phosphor(p)-AKT,however,knockdown of FBP1 led to the opposite.Furthermore,overexpression of MDM2 potently reversed FBP1-induced proliferation inhibition and apoptosis,while Nutlin-3(an MDM2 inhibitor)reversed the change trends induced by FBP1 knockdown in the aforementioned events.Lastly,but not least importantly,our data elucidated that MDM2 binds directly to FBP1 and promotes FBP1 ubiquitination.In conclusion,our data firstly suggested the involvement of FBP1 and its association with MDM2 in HCC cell growth.MDM2-FBP1-regulated HCC cell growth and the activation of AKT were mediated,at least in part,through FBP1 degradation.展开更多
Introduction:Fructose-1,6-bisphosphatase (FBPase) deficiency is a rare inherited disorder in gluconeogenesis,characterized by hypoglycemia,ketonuria,metabolic acidosis and convulsions.Case presentation:We describe two...Introduction:Fructose-1,6-bisphosphatase (FBPase) deficiency is a rare inherited disorder in gluconeogenesis,characterized by hypoglycemia,ketonuria,metabolic acidosis and convulsions.Case presentation:We describe two brothers with FBPase deficiency.The proband developed severe hypoglycemia and progressed to status epilepticus,and the brother showed slightly hypoglycemia with a good prognosis.Whole exome sequencing (WES) identified compound heterozygous variants [c.333+1333+2delinsTC and c.490G>A (p.Gly164Ser)] in fructose-1,6-bisphosphatase 1 gene in the two brothers,which were inherited from the father and the mother,respectively.Conclusion:Genetic analysis provided a solid basis for a definite diagnosis and the determination of precision therapies for the patient.展开更多
目的:研究活血化瘀治法(the medicine group of promoting blood circulation and removing stasis,PBCRS)对7,12-二甲基苯蒽[7,12-dimethylbenz(a) anthracene,DMBA]诱发性大鼠乳腺癌的抑制作用,及运用转录组测序技术(RNA sequencing,R...目的:研究活血化瘀治法(the medicine group of promoting blood circulation and removing stasis,PBCRS)对7,12-二甲基苯蒽[7,12-dimethylbenz(a) anthracene,DMBA]诱发性大鼠乳腺癌的抑制作用,及运用转录组测序技术(RNA sequencing,RNA-seq)和信号网络分析(ingenuity pathway analysis,IPA)软件筛选关键基因和验证。方法:将96只SD大鼠随机分为空白组,DMBA模型组,枸橼酸他莫昔芬(Tamoxifen,TAM)组(1.9 mg·kg^(-1)·d-1),PBCRS高、中、低剂量组(12.96,6.48,3.24 g·kg^(-1)·d-1)。药物干预1周后,除空白组外,采用DMBA诱导大鼠乳腺癌模型(灌服2次,间隔1周,剂量100 mg·kg^(-1))。给药10周后,观察大鼠荷瘤重、荷瘤体积变化,用总RNA提取试剂盒提取总RNA,从DMBA模型组和PBCRS中剂量组各取3个RNA样品进行基因检测,筛选出关键差异基因,用实时荧光定量PCR(Real-time PCR)验证关键基因。结果:与DMBA模型组比较,PBCRS中剂量组荷瘤重和荷瘤体积均明显降低(P<0.05),PBCRS高、低剂量组荷瘤重和荷瘤体积虽有降低,但与DMBA模型组比较差异均无统计学意义;基于RNA-seq技术及IPA分析软件筛选出果糖1,6二磷酸酶1 (fructose-1,6-bisphosphatase1,FBP1),浦肯野细胞蛋白4(purkinje cell protein 4,PCP4),肌红蛋白(myoglobin,MB) 3个关键基因;与DMBA模型组比较,PBCRS高、中剂量组FBP1 mRNA表达均明显升高(P<0.05),PBCRS低剂量组FBP1 mRNA表达升高,但差异无统计学意义,PBCRS高、中、低剂量组PCP4 mRNA的表达增高,但无统计学差异,PBCRS低剂量组MB mRNA的表达虽降低,差异无统计学意义。结论:PBCRS可能通过干预乳腺癌组织中FBP1的表达,抑制乳腺癌的发生。展开更多
基金The study was supported by the Grant QN201706 from Young Talent Training Project of Health Development Planning Commission of Changzhou City,and Grant CE20205034 from Science and Technology Supporting Project of Changzhou City.
文摘To study the roles and association of murine double minute gene 2(MDM2)and fructose-1,6-biphosphatase(FBP1)in human hepatocellular carcinoma(HCC),growth response of human HCC cells was assessed using proliferation and apoptosis assay.Pro-survival AKT signaling associated proteins(p-AKT,survivin and cleaved caspase 3)were assessed using western blotting.The correlation between MDM2 and FBP1 was assessed using coimmunoprecipitation combined with ubiquitination assay.Our data suggested that low expression of FBP1 was correlated with high levels of MDM2 in HCC cell lines(Huh7 and Hep3B).Overexpression of FBP1 resulted in antiproliferation,pro-apoptosis,the up-regulation of cleaved caspase 3 while the downregulation of survivin and phosphor(p)-AKT,however,knockdown of FBP1 led to the opposite.Furthermore,overexpression of MDM2 potently reversed FBP1-induced proliferation inhibition and apoptosis,while Nutlin-3(an MDM2 inhibitor)reversed the change trends induced by FBP1 knockdown in the aforementioned events.Lastly,but not least importantly,our data elucidated that MDM2 binds directly to FBP1 and promotes FBP1 ubiquitination.In conclusion,our data firstly suggested the involvement of FBP1 and its association with MDM2 in HCC cell growth.MDM2-FBP1-regulated HCC cell growth and the activation of AKT were mediated,at least in part,through FBP1 degradation.
基金grants from the National Natural Science Foundation of China(81701125)the Foundation of Henan Scientific committee(182102310418)+1 种基金the Foundation of Henan Provincial Department of health(2018020603)the Educational commission of Henan province of China(18A310029).
文摘Introduction:Fructose-1,6-bisphosphatase (FBPase) deficiency is a rare inherited disorder in gluconeogenesis,characterized by hypoglycemia,ketonuria,metabolic acidosis and convulsions.Case presentation:We describe two brothers with FBPase deficiency.The proband developed severe hypoglycemia and progressed to status epilepticus,and the brother showed slightly hypoglycemia with a good prognosis.Whole exome sequencing (WES) identified compound heterozygous variants [c.333+1333+2delinsTC and c.490G>A (p.Gly164Ser)] in fructose-1,6-bisphosphatase 1 gene in the two brothers,which were inherited from the father and the mother,respectively.Conclusion:Genetic analysis provided a solid basis for a definite diagnosis and the determination of precision therapies for the patient.
