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
Background:Inhibitors of B-cell CLL/Iymphoma 2(Bcl-2)family proteins have shown hope as antitumor drugs.While the notion that it is efficient to coordinate,balance,and neutralize both arms of the anti-apoptotic Bcl-2 ...Background:Inhibitors of B-cell CLL/Iymphoma 2(Bcl-2)family proteins have shown hope as antitumor drugs.While the notion that it is efficient to coordinate,balance,and neutralize both arms of the anti-apoptotic Bcl-2 family has been validated in many cancer cells,the weights of the two arms contributing to apoptosis inhibition have not been explored.This study analyzed the best combination ratio for different Bcl-2 selective inhibitors.Methods:We used a previously established mathematical model to study the weights of Bcl-2(representing both Bcl-2 and BcI-xL in this study)and myeloid cell leukemia-1(Mcl-1).Correlation and single-parameter sensitivity analysis were used to find the major molecular determinants for Bcl-2 and Mcl-1 dependency,as well as their weights.Biological experiments were used to verify the mathematical model.Results:Bcl-2 protein level and Mcl-1 protein level,production,and degradation rates were the major molecular determinants for Bcl-2 and Mcl-1 dependency.The model gained agreement with the experimental assays for ABT-737/A-1210477 and ABT-737/compound 5 combination effect in MCF-7 and MDA-MB-231.Two sets of equations composed of Bcl-2 and Mcl-1 levels were obtained to predict the best combination ratio for Bcl-2 inhibitors with Mcl-1 inhibitors that stabilize and downregulate Mcl-1,respectively.Conclusions:The two sets of equations can be used as tools to bypass time-consuming and laborious experimental screening to predict the best drug combination ratio for treatment.展开更多
Biliary tract cancers(BTC)are frequently identified at late stages and have a poor prognosis due to limited systemic treatment regimens.For more than a decade,the combination of gemcitabine and cis-platin has served a...Biliary tract cancers(BTC)are frequently identified at late stages and have a poor prognosis due to limited systemic treatment regimens.For more than a decade,the combination of gemcitabine and cis-platin has served as the first-line standard treatment.There are few choices for second-line chemo-therapy.Targeted treatment with fibroblast growth factor receptor 2 inhibitors,neurotrophic tyrosine receptor kinase inhibitors,and isocitrate dehydrogenase 1 inhibitors has had important results.Immune checkpoint inhibitors(ICI)such as pembrolizumab are only used in first-line treatment for microsatellite instability high patients.The TOPAZ-1 trial's outcome is encouraging,and there are several trials underway that might soon put targeted treatment and ICI combos into first-line options.Newer targets and agents for existing goals are being studied,which may represent a paradigm shift in BTC management.Due to a scarcity of targetable mutations and the higher toxicity profile of the current medications,the new category of drugs may occupy a significant role in BTC therapies.展开更多
The recent article by Zhou et al was highly interesting and thought provoking. The authors have clearly shown that triptolide administration is associated with upregulation of the Bax gene, resulting in an attenuating...The recent article by Zhou et al was highly interesting and thought provoking. The authors have clearly shown that triptolide administration is associated with upregulation of the Bax gene, resulting in an attenuating effect on cell growth in gastrointestinal malignancies such as pancreatic carcinomas. The article by Zhou et al is all the more important because it highlights the rapidly increasing role of triplodide in the management of systemic malignancies. For instance, triptolide acts on the PI3K/Akt/NF-κB pathway, thereby enhancing apoptosis secondary to the administration of bortezomib in multiple myeloma cells. Similar synergisms are seen when triptolide is administered along with 5-fluoruracil for the management of colonic carcinomas. Similarly, triptolide causes down-regulation of the Bcl-2 gene, resulting in control of cell growth in tumors, such as glioblastoma multiformes.展开更多
Cancer is cell fleeing from death by blocking the intrinsic and extrinsic programs of cell death. The six elements that shut down those programs are: muc-1, muc-4, muc-16, Bcl-2, MMPs and decoy R3. The nuclear factor-...Cancer is cell fleeing from death by blocking the intrinsic and extrinsic programs of cell death. The six elements that shut down those programs are: muc-1, muc-4, muc-16, Bcl-2, MMPs and decoy R3. The nuclear factor-kappa B (NF-Kb) stimulates the expression of genes responsible for the production of those elements, which are used by cells to block those two programs. In other words, the nuclear factor kappa B (NF-KB) is responsible for blocking both programs. Therefore, the nuclear factor kappa B (NF-Kb) is the true cause of cancer.展开更多
OBJECTIVE:To investigate the mechanism underlying the anticancer effect of Artemisia species through the inhibition of cell growth and induction of apoptosis in breast carcinoma cells.METHODS:To evaluate the anticance...OBJECTIVE:To investigate the mechanism underlying the anticancer effect of Artemisia species through the inhibition of cell growth and induction of apoptosis in breast carcinoma cells.METHODS:To evaluate the anticancer activity of methanol extracts of eight Artemisia species(Artemisia stolonifera,Artemisia selengensis,Artemisia japonica,Artemisia Montana,Artemisia capillaris,Artemisia sylvatica,Artemisia keiskeana,and Artemisia scoparia),we first investigated the proliferation of estrogen receptor(ER)-positive MCF-7breast carcinoma cells exposed to 5 or 200 g/mL for72 h.Apoptosis induction was assessed by an Annexin V binding assay in cells exposed to extracts at a high concentration(200 g/mL).To verify the mechanism of apoptosis,ER expression and its related signaling was investigated using an immunoblot assay under the same conditions.RESULTS:MCF-7 cells showed the strongest antiproliferative response to the tested extracts.Howev-er,a biphasic effect was observed:the extracts inhibited proliferation at high concentrations whereas they stimulated it at low ones.ER expression was similarly modulated by the extracts.However,all of the extracts induced apoptosis at a high concentration(200 g/mL).Compared to the control level,exposure to the extracts resulted in a remarkable increase in the shift of cell populations.CONCLUSION:The present study suggests that the tested Artemisia species exerted their anticancer effects through the induction of apoptosis via an ER-related pathway.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.
基金This research was supported by the National Natural Science Foundation of China(81430083,81903462 and 82073703)the China Postdoctoral Science Foundation(2018M641694)the Fundamental Research Funds for the Central Universities(DUT20LK28 and DUT20YG133).
文摘Background:Inhibitors of B-cell CLL/Iymphoma 2(Bcl-2)family proteins have shown hope as antitumor drugs.While the notion that it is efficient to coordinate,balance,and neutralize both arms of the anti-apoptotic Bcl-2 family has been validated in many cancer cells,the weights of the two arms contributing to apoptosis inhibition have not been explored.This study analyzed the best combination ratio for different Bcl-2 selective inhibitors.Methods:We used a previously established mathematical model to study the weights of Bcl-2(representing both Bcl-2 and BcI-xL in this study)and myeloid cell leukemia-1(Mcl-1).Correlation and single-parameter sensitivity analysis were used to find the major molecular determinants for Bcl-2 and Mcl-1 dependency,as well as their weights.Biological experiments were used to verify the mathematical model.Results:Bcl-2 protein level and Mcl-1 protein level,production,and degradation rates were the major molecular determinants for Bcl-2 and Mcl-1 dependency.The model gained agreement with the experimental assays for ABT-737/A-1210477 and ABT-737/compound 5 combination effect in MCF-7 and MDA-MB-231.Two sets of equations composed of Bcl-2 and Mcl-1 levels were obtained to predict the best combination ratio for Bcl-2 inhibitors with Mcl-1 inhibitors that stabilize and downregulate Mcl-1,respectively.Conclusions:The two sets of equations can be used as tools to bypass time-consuming and laborious experimental screening to predict the best drug combination ratio for treatment.
文摘Biliary tract cancers(BTC)are frequently identified at late stages and have a poor prognosis due to limited systemic treatment regimens.For more than a decade,the combination of gemcitabine and cis-platin has served as the first-line standard treatment.There are few choices for second-line chemo-therapy.Targeted treatment with fibroblast growth factor receptor 2 inhibitors,neurotrophic tyrosine receptor kinase inhibitors,and isocitrate dehydrogenase 1 inhibitors has had important results.Immune checkpoint inhibitors(ICI)such as pembrolizumab are only used in first-line treatment for microsatellite instability high patients.The TOPAZ-1 trial's outcome is encouraging,and there are several trials underway that might soon put targeted treatment and ICI combos into first-line options.Newer targets and agents for existing goals are being studied,which may represent a paradigm shift in BTC management.Due to a scarcity of targetable mutations and the higher toxicity profile of the current medications,the new category of drugs may occupy a significant role in BTC therapies.
文摘The recent article by Zhou et al was highly interesting and thought provoking. The authors have clearly shown that triptolide administration is associated with upregulation of the Bax gene, resulting in an attenuating effect on cell growth in gastrointestinal malignancies such as pancreatic carcinomas. The article by Zhou et al is all the more important because it highlights the rapidly increasing role of triplodide in the management of systemic malignancies. For instance, triptolide acts on the PI3K/Akt/NF-κB pathway, thereby enhancing apoptosis secondary to the administration of bortezomib in multiple myeloma cells. Similar synergisms are seen when triptolide is administered along with 5-fluoruracil for the management of colonic carcinomas. Similarly, triptolide causes down-regulation of the Bcl-2 gene, resulting in control of cell growth in tumors, such as glioblastoma multiformes.
文摘Cancer is cell fleeing from death by blocking the intrinsic and extrinsic programs of cell death. The six elements that shut down those programs are: muc-1, muc-4, muc-16, Bcl-2, MMPs and decoy R3. The nuclear factor-kappa B (NF-Kb) stimulates the expression of genes responsible for the production of those elements, which are used by cells to block those two programs. In other words, the nuclear factor kappa B (NF-KB) is responsible for blocking both programs. Therefore, the nuclear factor kappa B (NF-Kb) is the true cause of cancer.
基金Supported by Priority Research Centers Program through the National Research Foundation of Korea(NRF)funded by the Ministry of Education,Science and Technology(NRF-2009-0094017 and NRF-2011-0017017)
文摘OBJECTIVE:To investigate the mechanism underlying the anticancer effect of Artemisia species through the inhibition of cell growth and induction of apoptosis in breast carcinoma cells.METHODS:To evaluate the anticancer activity of methanol extracts of eight Artemisia species(Artemisia stolonifera,Artemisia selengensis,Artemisia japonica,Artemisia Montana,Artemisia capillaris,Artemisia sylvatica,Artemisia keiskeana,and Artemisia scoparia),we first investigated the proliferation of estrogen receptor(ER)-positive MCF-7breast carcinoma cells exposed to 5 or 200 g/mL for72 h.Apoptosis induction was assessed by an Annexin V binding assay in cells exposed to extracts at a high concentration(200 g/mL).To verify the mechanism of apoptosis,ER expression and its related signaling was investigated using an immunoblot assay under the same conditions.RESULTS:MCF-7 cells showed the strongest antiproliferative response to the tested extracts.Howev-er,a biphasic effect was observed:the extracts inhibited proliferation at high concentrations whereas they stimulated it at low ones.ER expression was similarly modulated by the extracts.However,all of the extracts induced apoptosis at a high concentration(200 g/mL).Compared to the control level,exposure to the extracts resulted in a remarkable increase in the shift of cell populations.CONCLUSION:The present study suggests that the tested Artemisia species exerted their anticancer effects through the induction of apoptosis via an ER-related pathway.
