Objective:Mcl-1 overexpression confers acquired resistance to Bcl-2 inhibitors in non-small cell lung cancer(NSCLC),but no direct Mcl-1 inhibitor is currently available for clinical use.Thus,novel therapeutic strategi...Objective:Mcl-1 overexpression confers acquired resistance to Bcl-2 inhibitors in non-small cell lung cancer(NSCLC),but no direct Mcl-1 inhibitor is currently available for clinical use.Thus,novel therapeutic strategies are urgently needed to target Mcl-1 and sensitize the anti-NSCLC activity of Bcl-2 inhibitors.Methods:Cell proliferation was measured using sulforhodamine B and colony formation assays,and apoptosis was detected with Annexin V-FITC staining.Gene expression was manipulated using siRNAs and plasmids.Real-time PCR and Western blot were used to measure mRNA and protein levels.Immunoprecipitation and immunofluorescence were used to analyze co-localization o f dual specificity tyrosine-phosphorylation-regulated kinase 1A(DYRK1A)and Mcl-1.Results:Suppression of DYRK1A resulted in reduced Mcl-1 expression in NSCLC cells,whereas overexpression of DYRK1A significantly increased Mcl-1 expression.Suppression of DYRK1A did not alter Mcl-1 mRNA levels,but did result in an accelerated degradation o f Mcl-1 protein in NSCLC cells.Furthermore,DYRK1A mediated proteasome-dependent degradation o f Mcl-1 in NSCLC cells,and DYRK1A co-localized with Mcl-1 in NSCLC cells and was co-expressed with Mcl-1 in tumor samples from lung cancer patients,suggesting that Mcl-1 may be a novel DYRK1A substrate.We showed that combined therapy with harmine and Bcl-2 antagonists significantly inhibited cell proliferation and induced apoptosis in NSCLC cell lines as well as primary NSCLC cells.Conclusions:Mcl-1 is a novel DYRK1A substrate,and the role of DYRK1A in promoting Mcl-1 stability makes it an attractive target for decreasing Bcl-2 inhibitor resistance.展开更多
In the published paper1,errors appeared in Figure 1D on page 391.In Figure 1D,an incorrect gel image was shown for Bcl-xL in NCI-H1299 cells.Figure 1D has been updated to correct the mistake above.The errors do not af...In the published paper1,errors appeared in Figure 1D on page 391.In Figure 1D,an incorrect gel image was shown for Bcl-xL in NCI-H1299 cells.Figure 1D has been updated to correct the mistake above.The errors do not affect the conclusions of this article.We apologize for the errors and for any confusion that they may have caused.展开更多
基金the National Natural Science Foundation of China(Grant Nos.81702887,81272473)the Key Laboratory of Clinical Cancer Pharmacology and Toxicology Research of Zhejiang Province(Grant No.2020E10021)+6 种基金the Key Medical Disciplines of Zhejiang Province(Grant No.2018-2-3)the Hangzhou Major Science and Technology Project(Grant No.20172016A01)the Zhejiang Provincial Foundation of Natural Science(Grant No.LY19H310004)the Scientific and Technological Developing Scheme of Hangzhou City(Grant No.20191203B49)the Science Research Foundation of Zhejiang Health Bureau(Grant No.2020RC026)the High-level Talents Coming Back from Abroad Innovation,the Entrepreneurship Program in Hangzhouand the Teachers Research Fund of Zhejiang University City College(Grant No.J-19006).
文摘Objective:Mcl-1 overexpression confers acquired resistance to Bcl-2 inhibitors in non-small cell lung cancer(NSCLC),but no direct Mcl-1 inhibitor is currently available for clinical use.Thus,novel therapeutic strategies are urgently needed to target Mcl-1 and sensitize the anti-NSCLC activity of Bcl-2 inhibitors.Methods:Cell proliferation was measured using sulforhodamine B and colony formation assays,and apoptosis was detected with Annexin V-FITC staining.Gene expression was manipulated using siRNAs and plasmids.Real-time PCR and Western blot were used to measure mRNA and protein levels.Immunoprecipitation and immunofluorescence were used to analyze co-localization o f dual specificity tyrosine-phosphorylation-regulated kinase 1A(DYRK1A)and Mcl-1.Results:Suppression of DYRK1A resulted in reduced Mcl-1 expression in NSCLC cells,whereas overexpression of DYRK1A significantly increased Mcl-1 expression.Suppression of DYRK1A did not alter Mcl-1 mRNA levels,but did result in an accelerated degradation o f Mcl-1 protein in NSCLC cells.Furthermore,DYRK1A mediated proteasome-dependent degradation o f Mcl-1 in NSCLC cells,and DYRK1A co-localized with Mcl-1 in NSCLC cells and was co-expressed with Mcl-1 in tumor samples from lung cancer patients,suggesting that Mcl-1 may be a novel DYRK1A substrate.We showed that combined therapy with harmine and Bcl-2 antagonists significantly inhibited cell proliferation and induced apoptosis in NSCLC cell lines as well as primary NSCLC cells.Conclusions:Mcl-1 is a novel DYRK1A substrate,and the role of DYRK1A in promoting Mcl-1 stability makes it an attractive target for decreasing Bcl-2 inhibitor resistance.
文摘In the published paper1,errors appeared in Figure 1D on page 391.In Figure 1D,an incorrect gel image was shown for Bcl-xL in NCI-H1299 cells.Figure 1D has been updated to correct the mistake above.The errors do not affect the conclusions of this article.We apologize for the errors and for any confusion that they may have caused.
