Objective:Pancreatic ductal adenocarcinoma(PDAC)is a highly malignant gastrointestinal cancer with a 5-year survival rate of only 9%.Of PDAC patients,15%-20%are eligible for radical surgery.Gemcitabine is an important...Objective:Pancreatic ductal adenocarcinoma(PDAC)is a highly malignant gastrointestinal cancer with a 5-year survival rate of only 9%.Of PDAC patients,15%-20%are eligible for radical surgery.Gemcitabine is an important chemotherapeutic agent for patients with PDAC;however,the efficacy of gemcitabine is limited due to resistance.Therefore,reducing gemcitabine resistance is essential for improving survival of patients with PDAC.Identifying the key target that determines gemcitabine resistance in PDAC and reversing gemcitabine resistance using target inhibitors in combination with gemcitabine are crucial steps in the quest to improve survival prognosis in patients with PDAC.Methods:We constructed a human genome-wide CRISPRa/dCas 9 overexpression library in PDAC cell lines to screen key targets of drug resistance based on sgRNA abundance and enrichment.Then,co-IP,ChIP,ChIP-seq,transcriptome sequencing,and qPCR were used to determine the specific mechanism by which phospholipase D1(PLD1)confers resistance to gemcitabine.Results:PLD1 combines with nucleophosmin 1(NPM1)and triggers NPM1 nuclear translocation,where NPM1 acts as a transcription factor to upregulate interleukin 7 receptor(IL7R)expression.Upon interleukin 7(IL-7)binding,IL7R activates the JAK1/STAT5 signaling pathway to increase the expression of the anti-apoptotic protein,BCL-2,and induce gemcitabine resistance.The PLD1 inhibitor,Vu0155069,targets PLD1 to induce apoptosis in gemcitabine-resistant PDAC cells.Conclusions:PLD1 is an enzyme that has a critical role in PDAC-associated gemcitabine resistance through a non-enzymatic interaction with NPM1,further promoting the downstream JAK1/STAT5/Bcl-2 pathway.Inhibiting any of the participants of this pathway can increase gemcitabine sensitivity.展开更多
AIM To determine the prevalence and diagnostic value of autoantibodies inα-fetoprotein(AFP)-negative hepatocellular carcinoma(HCC).METHODS Fifty-six serum samples from AFP-negative HCC cases,86 from AFP-positive HCC ...AIM To determine the prevalence and diagnostic value of autoantibodies inα-fetoprotein(AFP)-negative hepatocellular carcinoma(HCC).METHODS Fifty-six serum samples from AFP-negative HCC cases,86 from AFP-positive HCC cases,168 from chronic liver disease cases,and 59 from normal human controls were included in this study.Autoantibodies to nucleophosmin(NPM)1,14-3-3zeta and mouse double minute 2 homolog(MDM2)proteins in AFP-negative HCC serum were evaluated by enzymelinked im munosorbent assay.Partially positive sera were further evaluated by western blotting.Immunohistochemistry was used to detect the expression of three tumor-associated antigens(TAAs)in AFP-negative HCC and normal control tissues.RESULTS The frequency of autoantibodies to the three TAAs in AFP-negative HCC sera was 21.4%,19.6%and 19.6%,which was significantly higher than in the chronic liver disease cases and normal human controls(P<0.01)as well as AFP-positive HCC cases.The sensitivity of the three autoantibodies for diagnosis of AFP-negative HCC ranged from 19.6%to 21.4%,and the specificity was approximately 95%.When the three autoantibodies were combined,the sensitivity reached 30.4%and the specificity reached 91.6%.CONCLUSION Autoantibodies to NPM1,14-3-3zeta and MDM2 may be useful biomarkers for immunodiagnosis of AFP-negative HCC.展开更多
Mutations of fms-like tyrosine kinase 3(FLT3) and nucleophosmin(NPM1) exon 12 genes are the most common abnormalities in adult acute myeloid leukemia(AML) with normal cytogenetics.To assess the prognostic impact of th...Mutations of fms-like tyrosine kinase 3(FLT3) and nucleophosmin(NPM1) exon 12 genes are the most common abnormalities in adult acute myeloid leukemia(AML) with normal cytogenetics.To assess the prognostic impact of the two gene mutations in Chinese AML patients,we used multiplex polymerase chain reaction(PCR) and capillary electrophoresis to screen 76 AML patients with normal cytogenetics for mutations in FLT3 internal tandem duplication(FLT3/ITD) and exon 12 of the NPM1 gene.FLT3/ITD mutation was detected in 15(19.7%) of 76 subjects,and NPM1 mutation in 20(26.3%) subjects.Seven(9.2%) cases were positive for both FLT3/ITD and NPM1 mutations.Significantly more FLT3/ITD aberration was detected in subjects with French-American-British(FAB) M1(42.8%).NPM1 mutation was frequently detected in subjects with M5(47.1%) and infrequently in subjects with M2(11.1%).FLT3 and NPM1 mutations were significantly associated with a higher white blood cell count in peripheral blood and a lower CD34 antigen expression,but not age,sex,or platelet count.Statistical analysis revealed that the FLT3/ITD-positive group had a lower complete remission(CR) rate(53.3% vs.83.6%).Survival analysis showed that the FLT3/ITD-positive/NPM1 mutation-negative group had worse overall survival(OS) and relapse-free survival(RFS).The FLT3/ITD-positive/NPM1 mutation-positive group showed a trend towards favorable survival compared with the FLT3/ITD-positive/NPM1 mutation-negative group(P=0.069).Our results indicate that the FLT3/ITD mutation might be a prognostic factor for an unfavorable outcome in Chinese AML subjects with normal cytogenetics,while NPM1 mutation may be a favorable prognostic factor for OS and RFS in the presence of FLT3/ITD.展开更多
基金supported by the National Key Research and Development Program of China(Grant No.2021YFA1201100)the National Natural Science Foundation of China(Grant Nos.82103006,82030092,81720108028,82072657,82072716,82103003,82173295,81871968,81871978,82072691,and 82103222)+1 种基金the Tianjin Hygiene Healthy Science and Technology Project(Grant No.TJWJ2022MS007)the Science&Technology Development Fund of Tianjin Education Commission for Higher Education(Grant No.2020KJ141).
文摘Objective:Pancreatic ductal adenocarcinoma(PDAC)is a highly malignant gastrointestinal cancer with a 5-year survival rate of only 9%.Of PDAC patients,15%-20%are eligible for radical surgery.Gemcitabine is an important chemotherapeutic agent for patients with PDAC;however,the efficacy of gemcitabine is limited due to resistance.Therefore,reducing gemcitabine resistance is essential for improving survival of patients with PDAC.Identifying the key target that determines gemcitabine resistance in PDAC and reversing gemcitabine resistance using target inhibitors in combination with gemcitabine are crucial steps in the quest to improve survival prognosis in patients with PDAC.Methods:We constructed a human genome-wide CRISPRa/dCas 9 overexpression library in PDAC cell lines to screen key targets of drug resistance based on sgRNA abundance and enrichment.Then,co-IP,ChIP,ChIP-seq,transcriptome sequencing,and qPCR were used to determine the specific mechanism by which phospholipase D1(PLD1)confers resistance to gemcitabine.Results:PLD1 combines with nucleophosmin 1(NPM1)and triggers NPM1 nuclear translocation,where NPM1 acts as a transcription factor to upregulate interleukin 7 receptor(IL7R)expression.Upon interleukin 7(IL-7)binding,IL7R activates the JAK1/STAT5 signaling pathway to increase the expression of the anti-apoptotic protein,BCL-2,and induce gemcitabine resistance.The PLD1 inhibitor,Vu0155069,targets PLD1 to induce apoptosis in gemcitabine-resistant PDAC cells.Conclusions:PLD1 is an enzyme that has a critical role in PDAC-associated gemcitabine resistance through a non-enzymatic interaction with NPM1,further promoting the downstream JAK1/STAT5/Bcl-2 pathway.Inhibiting any of the participants of this pathway can increase gemcitabine sensitivity.
文摘目的:观察核仁磷酸蛋白(nucleophosmin 1,NPM1)A型突变体过表达对TGF-β1诱导的K562细胞增殖及AKT磷酸化的影响。方法:腺病毒载体(Ad5-NPM1)转染髓系白血病K562细胞建立过表达NPM1蛋白细胞株;应用Western blot法及ELISA法分别检测细胞NPM1蛋白表达及上清液含量,Westernblot法检测K562细胞NPM1、AKT和P-AKT蛋白表达,MTT法检测细胞增殖。结果:K562细胞NPM1蛋白表达水平呈Ad5-NPM1转染复数[(multiplicity of infection,MOI):30-200]依赖性增加,与空载组(Ad5-vector-100)相比,Ad5-NPM1-30、Ad5-NPM1-100组K562细胞及细胞上清液NPM1蛋白水平均显著增高(P<0.01)。TGF-β1(10ng/mL)能够诱导K562细胞AKT蛋白磷酸化,但对总AKT水平无明显影响。TGF-β1(10ng/mL)+Ad5-NPM1-100组K562细胞P-AKT水平显著高于TGF-β1处理组(P<0.05),各组总AKT水平无显著差别(P>0.05)。与空白对照组(CT)相比,TGF-β1(10ng/mL)处理可促进K562细胞增殖;但TGF-β1(10ng/mL)+Ad5-NPM1-100组细胞的增殖水平明显高于TGF-β1处理组(P<0.01)。结论:NPM1可促进TGF-β1诱导的K562细胞增殖。NPM1促进TGF-β1诱导的K562细胞可能与AKT磷酸化有关。
基金Supported by Clinical Research Cooperation Fund of the Capital Medical University,No.15JL67Project of Science and Technology Development Plan of Beijing Municipal Education Commission,No.KM201610025021+2 种基金High-Tech Personnel Training Program of Beijing Health System,No.2015-3-104Beijing Municipal Science and Technology Commission,No.Z151100004015066Shaanxi Science and Technology Coordination and Innovation Project,No.2016KTZDSF02-02
文摘AIM To determine the prevalence and diagnostic value of autoantibodies inα-fetoprotein(AFP)-negative hepatocellular carcinoma(HCC).METHODS Fifty-six serum samples from AFP-negative HCC cases,86 from AFP-positive HCC cases,168 from chronic liver disease cases,and 59 from normal human controls were included in this study.Autoantibodies to nucleophosmin(NPM)1,14-3-3zeta and mouse double minute 2 homolog(MDM2)proteins in AFP-negative HCC serum were evaluated by enzymelinked im munosorbent assay.Partially positive sera were further evaluated by western blotting.Immunohistochemistry was used to detect the expression of three tumor-associated antigens(TAAs)in AFP-negative HCC and normal control tissues.RESULTS The frequency of autoantibodies to the three TAAs in AFP-negative HCC sera was 21.4%,19.6%and 19.6%,which was significantly higher than in the chronic liver disease cases and normal human controls(P<0.01)as well as AFP-positive HCC cases.The sensitivity of the three autoantibodies for diagnosis of AFP-negative HCC ranged from 19.6%to 21.4%,and the specificity was approximately 95%.When the three autoantibodies were combined,the sensitivity reached 30.4%and the specificity reached 91.6%.CONCLUSION Autoantibodies to NPM1,14-3-3zeta and MDM2 may be useful biomarkers for immunodiagnosis of AFP-negative HCC.
文摘Mutations of fms-like tyrosine kinase 3(FLT3) and nucleophosmin(NPM1) exon 12 genes are the most common abnormalities in adult acute myeloid leukemia(AML) with normal cytogenetics.To assess the prognostic impact of the two gene mutations in Chinese AML patients,we used multiplex polymerase chain reaction(PCR) and capillary electrophoresis to screen 76 AML patients with normal cytogenetics for mutations in FLT3 internal tandem duplication(FLT3/ITD) and exon 12 of the NPM1 gene.FLT3/ITD mutation was detected in 15(19.7%) of 76 subjects,and NPM1 mutation in 20(26.3%) subjects.Seven(9.2%) cases were positive for both FLT3/ITD and NPM1 mutations.Significantly more FLT3/ITD aberration was detected in subjects with French-American-British(FAB) M1(42.8%).NPM1 mutation was frequently detected in subjects with M5(47.1%) and infrequently in subjects with M2(11.1%).FLT3 and NPM1 mutations were significantly associated with a higher white blood cell count in peripheral blood and a lower CD34 antigen expression,but not age,sex,or platelet count.Statistical analysis revealed that the FLT3/ITD-positive group had a lower complete remission(CR) rate(53.3% vs.83.6%).Survival analysis showed that the FLT3/ITD-positive/NPM1 mutation-negative group had worse overall survival(OS) and relapse-free survival(RFS).The FLT3/ITD-positive/NPM1 mutation-positive group showed a trend towards favorable survival compared with the FLT3/ITD-positive/NPM1 mutation-negative group(P=0.069).Our results indicate that the FLT3/ITD mutation might be a prognostic factor for an unfavorable outcome in Chinese AML subjects with normal cytogenetics,while NPM1 mutation may be a favorable prognostic factor for OS and RFS in the presence of FLT3/ITD.
