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Leukemia Stem Cell-Released Microvesicles Promote the Survival and Migration of Myeloid Leukemia Cells and These Effects Can Be Inhibited by MicroRNA34a Overexpression.

Wang Y, Cheng Q, Liu J, Dong M - Stem Cells Int (2016)

Bottom Line: The effect of miR34a transfection on LSC proliferation and the effects of LMVs(miRCtrl) or LMVs(miR34a) on the proliferation, migration, and apoptosis of AML cells (after LSC depletion) were determined.Results showed that (1) LMVs(miRCtrl) promoted proliferation and migration and inhibited apoptosis of AML cells, which were associated with miR34a deficit; (2) transfection of miR34a mimic inhibited LSC proliferation and increased miR34a level in LMVs(miR34a); (3) LMVs(miR34a) produced opposite effects as compared with LMVs(miRCtrl), which were associated with the changes of caspase-3 and Tim-3 levels.In summary, LMVs support AML cell malignance and modulating miR34a could offer a new approach for the management of AML.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, The Affiliated Hospital of Guilin Medical College, Guilin, Guangxi 541001, China.

ABSTRACT
Leukemia stem cells (LSCs) play the major role in relapse of acute myeloid leukemia (AML). Recent evidence indicates that microvesicles (MVs) released from cancer stem cells can promote tumor growth and invasion. In this study, we investigated whether LSCs-released MVs (LMVs) can regulate the malignance of AML cells and whether overexpression of tumor suppressive microRNA (miR), miR34a, is able to interrupt this process. LSCs were transfected with miRNA control (miRCtrl) or miR34a mimic for producing LMVs, respectively, defined as LMVs(miRCtrl) and LMVs(miR34a). The effect of miR34a transfection on LSC proliferation and the effects of LMVs(miRCtrl) or LMVs(miR34a) on the proliferation, migration, and apoptosis of AML cells (after LSC depletion) were determined. The levels of miR34a targets, caspase-3 and T cell immunoglobulin mucin-3 (Tim-3), were analyzed. Results showed that (1) LMVs(miRCtrl) promoted proliferation and migration and inhibited apoptosis of AML cells, which were associated with miR34a deficit; (2) transfection of miR34a mimic inhibited LSC proliferation and increased miR34a level in LMVs(miR34a); (3) LMVs(miR34a) produced opposite effects as compared with LMVs(miRCtrl), which were associated with the changes of caspase-3 and Tim-3 levels. In summary, LMVs support AML cell malignance and modulating miR34a could offer a new approach for the management of AML.

No MeSH data available.


Related in: MedlinePlus

LSC characterization and the effect of miR34a overexpression on LSC proliferation. (a) Representative flow cytometric plots of CD34-PE versus CD38-FITC. Expressions of CD34 and CD38 in KG1a cells before sorting (left) and after sorting (right). (b) LSCs were subjected to vehicle (veh) treatment and transfected with miRCtrl or miR34a mimic as indicated and 2 days later the miR34a level was measured by qRT-PCR. (c) LSCs were subjected to treatment with veh or miRCtrl transfection or miR34a mimic transfection. After 2 days, the LSC proliferation was determined by MTT assay. Data represents mean ± SEM. ∗P < 0.05 versus veh; #P < 0.05 versus miRCtrl.
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fig1: LSC characterization and the effect of miR34a overexpression on LSC proliferation. (a) Representative flow cytometric plots of CD34-PE versus CD38-FITC. Expressions of CD34 and CD38 in KG1a cells before sorting (left) and after sorting (right). (b) LSCs were subjected to vehicle (veh) treatment and transfected with miRCtrl or miR34a mimic as indicated and 2 days later the miR34a level was measured by qRT-PCR. (c) LSCs were subjected to treatment with veh or miRCtrl transfection or miR34a mimic transfection. After 2 days, the LSC proliferation was determined by MTT assay. Data represents mean ± SEM. ∗P < 0.05 versus veh; #P < 0.05 versus miRCtrl.

Mentions: To determine the proportion of LSCs in KG1a cells and whether LSCs were successfully isolated from KG1a cells, KG1a cells and microbeads-isolated LSCs were stained with CD34-PE and CD38-FITC antibodies [18, 19] and analyzed by flow cytometry. As shown in Figure 1(a), the KG1a cells contained about 32.8% of CD34+CD38− cells. After microbead sorting procedure, more than 95% of the cells were CD34+CD38− and considered LSCs.


Leukemia Stem Cell-Released Microvesicles Promote the Survival and Migration of Myeloid Leukemia Cells and These Effects Can Be Inhibited by MicroRNA34a Overexpression.

Wang Y, Cheng Q, Liu J, Dong M - Stem Cells Int (2016)

LSC characterization and the effect of miR34a overexpression on LSC proliferation. (a) Representative flow cytometric plots of CD34-PE versus CD38-FITC. Expressions of CD34 and CD38 in KG1a cells before sorting (left) and after sorting (right). (b) LSCs were subjected to vehicle (veh) treatment and transfected with miRCtrl or miR34a mimic as indicated and 2 days later the miR34a level was measured by qRT-PCR. (c) LSCs were subjected to treatment with veh or miRCtrl transfection or miR34a mimic transfection. After 2 days, the LSC proliferation was determined by MTT assay. Data represents mean ± SEM. ∗P < 0.05 versus veh; #P < 0.05 versus miRCtrl.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4835649&req=5

fig1: LSC characterization and the effect of miR34a overexpression on LSC proliferation. (a) Representative flow cytometric plots of CD34-PE versus CD38-FITC. Expressions of CD34 and CD38 in KG1a cells before sorting (left) and after sorting (right). (b) LSCs were subjected to vehicle (veh) treatment and transfected with miRCtrl or miR34a mimic as indicated and 2 days later the miR34a level was measured by qRT-PCR. (c) LSCs were subjected to treatment with veh or miRCtrl transfection or miR34a mimic transfection. After 2 days, the LSC proliferation was determined by MTT assay. Data represents mean ± SEM. ∗P < 0.05 versus veh; #P < 0.05 versus miRCtrl.
Mentions: To determine the proportion of LSCs in KG1a cells and whether LSCs were successfully isolated from KG1a cells, KG1a cells and microbeads-isolated LSCs were stained with CD34-PE and CD38-FITC antibodies [18, 19] and analyzed by flow cytometry. As shown in Figure 1(a), the KG1a cells contained about 32.8% of CD34+CD38− cells. After microbead sorting procedure, more than 95% of the cells were CD34+CD38− and considered LSCs.

Bottom Line: The effect of miR34a transfection on LSC proliferation and the effects of LMVs(miRCtrl) or LMVs(miR34a) on the proliferation, migration, and apoptosis of AML cells (after LSC depletion) were determined.Results showed that (1) LMVs(miRCtrl) promoted proliferation and migration and inhibited apoptosis of AML cells, which were associated with miR34a deficit; (2) transfection of miR34a mimic inhibited LSC proliferation and increased miR34a level in LMVs(miR34a); (3) LMVs(miR34a) produced opposite effects as compared with LMVs(miRCtrl), which were associated with the changes of caspase-3 and Tim-3 levels.In summary, LMVs support AML cell malignance and modulating miR34a could offer a new approach for the management of AML.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, The Affiliated Hospital of Guilin Medical College, Guilin, Guangxi 541001, China.

ABSTRACT
Leukemia stem cells (LSCs) play the major role in relapse of acute myeloid leukemia (AML). Recent evidence indicates that microvesicles (MVs) released from cancer stem cells can promote tumor growth and invasion. In this study, we investigated whether LSCs-released MVs (LMVs) can regulate the malignance of AML cells and whether overexpression of tumor suppressive microRNA (miR), miR34a, is able to interrupt this process. LSCs were transfected with miRNA control (miRCtrl) or miR34a mimic for producing LMVs, respectively, defined as LMVs(miRCtrl) and LMVs(miR34a). The effect of miR34a transfection on LSC proliferation and the effects of LMVs(miRCtrl) or LMVs(miR34a) on the proliferation, migration, and apoptosis of AML cells (after LSC depletion) were determined. The levels of miR34a targets, caspase-3 and T cell immunoglobulin mucin-3 (Tim-3), were analyzed. Results showed that (1) LMVs(miRCtrl) promoted proliferation and migration and inhibited apoptosis of AML cells, which were associated with miR34a deficit; (2) transfection of miR34a mimic inhibited LSC proliferation and increased miR34a level in LMVs(miR34a); (3) LMVs(miR34a) produced opposite effects as compared with LMVs(miRCtrl), which were associated with the changes of caspase-3 and Tim-3 levels. In summary, LMVs support AML cell malignance and modulating miR34a could offer a new approach for the management of AML.

No MeSH data available.


Related in: MedlinePlus