As one of the hallmarks of cancer,metabolic reprogramming leads to cancer progression,and targeting glycolytic enzymes could be useful strategies for cancer therapy.By screening a small molecule library consisting of ...As one of the hallmarks of cancer,metabolic reprogramming leads to cancer progression,and targeting glycolytic enzymes could be useful strategies for cancer therapy.By screening a small molecule library consisting of 1320 FDA-approved drugs,we found that penfluridol,an antipsychotic drug used to treat schizophrenia,could inhibit glycolysis and induce apoptosis in esophageal squamous cell carcinoma(ESCC).Gene profiling and Ingenuity Pathway Analysis suggested the important role of AMPK in action mechanism of penfluridol.By using drug affinity responsive target stability(DARTS)technology and proteomics,we identified phosphofructokinase,liver type(PFKL),a key enzyme in glycolysis,as a direct target of penfluridol.Penfluridol could not exhibit its anticancer property in PFKL-deficient cancer cells,illustrating that PFKL is essential for the bioactivity of penfluridol.High PFKL expression is correlated with advanced stages and poor survival of ESCC patients,and silencing of PFKL significantly suppressed tumor growth.Mechanistically,direct binding of penfluridol and PFKL inhibits glucose consumption,lactate and ATP production,leads to nuclear translocation of FOXO3a and subsequent transcriptional activation of BIM in an AMPK-dependent manner.Taken together,PFKL is a potential prognostic biomarker and therapeutic target in ESCC,and penfluridol may be a new therapeutic option for management of this lethal disease.展开更多
目的建立人体全血中五氟利多浓度的液相色谱-质谱联用法(LC-MS/MS)分析方法。方法全血中五氟利多和利培酮(内标)经正己烷液-液提取后,采用Capcell Pak C18色谱柱(250mm×2.0mm5,μm)进行分离,流动相为乙腈:20mmol/L乙酸胺和0.1%甲...目的建立人体全血中五氟利多浓度的液相色谱-质谱联用法(LC-MS/MS)分析方法。方法全血中五氟利多和利培酮(内标)经正己烷液-液提取后,采用Capcell Pak C18色谱柱(250mm×2.0mm5,μm)进行分离,流动相为乙腈:20mmol/L乙酸胺和0.1%甲酸溶液(75∶25,V/V),流速为0.2mL/min,然后以MS/MS电喷雾正电离的多反应监测扫描方式(MRM)测定。用于定量分析的离子为m/z 524→109(五氟利多)和m/z 411→191(内标)。结果五氟利多的最低检测限为0.2ng/mL,在0.4～400ng/mL浓度范围内线性良好(r=0.9994),低、中、高浓度(1ng/mL、10ng/mL、100ng/mL)准确度分别为97%,108%和95%,日内和日间RSD均小于15%。结论该方法简便、快速、灵敏,适用于全血中五氟利多浓度的测定。展开更多
基金supported by National Natural Science Foundation of China(31961160727,81773085,and 81973339)National Key Research and Development Program of China(2017YFA0505100)Guangdong Natural Science Research Grant International joint project(2021A0505030035,China)。
文摘As one of the hallmarks of cancer,metabolic reprogramming leads to cancer progression,and targeting glycolytic enzymes could be useful strategies for cancer therapy.By screening a small molecule library consisting of 1320 FDA-approved drugs,we found that penfluridol,an antipsychotic drug used to treat schizophrenia,could inhibit glycolysis and induce apoptosis in esophageal squamous cell carcinoma(ESCC).Gene profiling and Ingenuity Pathway Analysis suggested the important role of AMPK in action mechanism of penfluridol.By using drug affinity responsive target stability(DARTS)technology and proteomics,we identified phosphofructokinase,liver type(PFKL),a key enzyme in glycolysis,as a direct target of penfluridol.Penfluridol could not exhibit its anticancer property in PFKL-deficient cancer cells,illustrating that PFKL is essential for the bioactivity of penfluridol.High PFKL expression is correlated with advanced stages and poor survival of ESCC patients,and silencing of PFKL significantly suppressed tumor growth.Mechanistically,direct binding of penfluridol and PFKL inhibits glucose consumption,lactate and ATP production,leads to nuclear translocation of FOXO3a and subsequent transcriptional activation of BIM in an AMPK-dependent manner.Taken together,PFKL is a potential prognostic biomarker and therapeutic target in ESCC,and penfluridol may be a new therapeutic option for management of this lethal disease.
文摘目的建立人体全血中五氟利多浓度的液相色谱-质谱联用法(LC-MS/MS)分析方法。方法全血中五氟利多和利培酮(内标)经正己烷液-液提取后,采用Capcell Pak C18色谱柱(250mm×2.0mm5,μm)进行分离,流动相为乙腈:20mmol/L乙酸胺和0.1%甲酸溶液(75∶25,V/V),流速为0.2mL/min,然后以MS/MS电喷雾正电离的多反应监测扫描方式(MRM)测定。用于定量分析的离子为m/z 524→109(五氟利多)和m/z 411→191(内标)。结果五氟利多的最低检测限为0.2ng/mL,在0.4～400ng/mL浓度范围内线性良好(r=0.9994),低、中、高浓度(1ng/mL、10ng/mL、100ng/mL)准确度分别为97%,108%和95%,日内和日间RSD均小于15%。结论该方法简便、快速、灵敏,适用于全血中五氟利多浓度的测定。
