Objective： To study the mechanisms in gambogic acid （GA） -induced JeKo-1 human Mantle Cell Lymphoma cell apoptosis in vitro. Methods： The proliferation of GA-treated JeKo-1 cells was measured by CCK-8 assay and Ki...Objective： To study the mechanisms in gambogic acid （GA） -induced JeKo-1 human Mantle Cell Lymphoma cell apoptosis in vitro. Methods： The proliferation of GA-treated JeKo-1 cells was measured by CCK-8 assay and Ki-67 immunocytochemical detection. Apopt0sis, cell cycle and mitochondrial membrane potential were measured by flow cytometric analysis. Caspase-3, -8 and -9 were detected by colorimetric assay. Bcl-2 and Bax were analyzed by Western blotting. Results： GA inhibited cell growth in a time- and dose- dependent manner. GA induces apoptosis in JeKo- 1 cells but not in normal bone marrow cells, which was involved in reducing the membrane potential of mitochondria, activating caspases-3, -8 and -9 and decreasing the ratio of Bd-2 and Bax without cell cycle arresting. Conclusions： GA induced apoptosis in human MCL JeKo-1 cells by regulating Bcl-2/Bax and activating caspase-3, -8 and -9 via mitochondrial pathway without affecting cell cycle.展开更多
Mantle cell lymphoma(MCL)is a B-cell malignancy with poor clinical outcome and undefined pathogenesis.Development of clinically relevant cellular models for MCL research is an urgent need.Our preliminary observations ...Mantle cell lymphoma(MCL)is a B-cell malignancy with poor clinical outcome and undefined pathogenesis.Development of clinically relevant cellular models for MCL research is an urgent need.Our preliminary observations lead the development of two novel hypotheses that we tested in this study:1.multicellular spheroid might be a unique growth mode of earlystage cells in MCL;2.MCL might be a polyclonal tumor.We made the following original observations that have not been reported:First,we have provided a new experiment method for enriching MCL early-stage cells and characterized the spheroid mode of growth as a unique feature of early-stage MCL cells in cell line as well as in clinical samples.Second,we have established a clinically relevant cellular model of MCL,the JeKo-1-spheroid cell line,that was highly enriched in early-stage sub-clones.JeKo-1-spheroid cells and the spheroid growing cells enriched from MCL patients exhibited comparably enhanced tumorigenic abilities and similar biological features.Third,Immunophenotypic analysis has revealed that MCL may be derived from precursor-B(pre-B),immature-B and mature-B cells,not only the mature-B cells as WHO classified in 2016.Fourth,MCL may be a polyclonal disease composed of CD19e/IgMe,CD19 e/IgMt,CD19t/IgMt three sub-clones,of which the CD19e/IgMt sub-clone might be the dominant sub-clone with the strongest tumorigenic ability.Fifth,CD19t/IgMe that differentiates MCL and normal B cells may represent a new marker for MCL early detection,minor residual disease monitoring after therapies and prognosis.展开更多
基金supported by a grant from the Key Project supported by medical science and technology development Foundation of Nanjing Department of Health (No. ZKX09016)
文摘Objective： To study the mechanisms in gambogic acid （GA） -induced JeKo-1 human Mantle Cell Lymphoma cell apoptosis in vitro. Methods： The proliferation of GA-treated JeKo-1 cells was measured by CCK-8 assay and Ki-67 immunocytochemical detection. Apopt0sis, cell cycle and mitochondrial membrane potential were measured by flow cytometric analysis. Caspase-3, -8 and -9 were detected by colorimetric assay. Bcl-2 and Bax were analyzed by Western blotting. Results： GA inhibited cell growth in a time- and dose- dependent manner. GA induces apoptosis in JeKo- 1 cells but not in normal bone marrow cells, which was involved in reducing the membrane potential of mitochondria, activating caspases-3, -8 and -9 and decreasing the ratio of Bd-2 and Bax without cell cycle arresting. Conclusions： GA induced apoptosis in human MCL JeKo-1 cells by regulating Bcl-2/Bax and activating caspase-3, -8 and -9 via mitochondrial pathway without affecting cell cycle.
基金We thank all the patients,healthy donors and their families,as well as The Affiliated Hospital of Southwest Medical University and Mianyang central hospital participating in this trial.We are thankful to the members in the Laboratory of Translational Cancer Stem Cell Research who are not listed in the authors.This work was funded by the National Natural Science Fund(Grant No.81272405)the Funds for Luzhou Medical College Applied Basic Research Plan(Grant No.2015-YJ122)+3 种基金the Key Research Project from Health and Family Planning Commission of Sichuan Province(Grant No.18ZD014)the National Natural Science Fund(Grant No.81450030)the Key Research Project of Sichuan Education Department(Grant No.14ZA0141)the Luzhou Science and Technology Project(Grant No.2014-S-47).
文摘Mantle cell lymphoma(MCL)is a B-cell malignancy with poor clinical outcome and undefined pathogenesis.Development of clinically relevant cellular models for MCL research is an urgent need.Our preliminary observations lead the development of two novel hypotheses that we tested in this study:1.multicellular spheroid might be a unique growth mode of earlystage cells in MCL;2.MCL might be a polyclonal tumor.We made the following original observations that have not been reported:First,we have provided a new experiment method for enriching MCL early-stage cells and characterized the spheroid mode of growth as a unique feature of early-stage MCL cells in cell line as well as in clinical samples.Second,we have established a clinically relevant cellular model of MCL,the JeKo-1-spheroid cell line,that was highly enriched in early-stage sub-clones.JeKo-1-spheroid cells and the spheroid growing cells enriched from MCL patients exhibited comparably enhanced tumorigenic abilities and similar biological features.Third,Immunophenotypic analysis has revealed that MCL may be derived from precursor-B(pre-B),immature-B and mature-B cells,not only the mature-B cells as WHO classified in 2016.Fourth,MCL may be a polyclonal disease composed of CD19e/IgMe,CD19 e/IgMt,CD19t/IgMt three sub-clones,of which the CD19e/IgMt sub-clone might be the dominant sub-clone with the strongest tumorigenic ability.Fifth,CD19t/IgMe that differentiates MCL and normal B cells may represent a new marker for MCL early detection,minor residual disease monitoring after therapies and prognosis.
