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In vivo and in vitro effects of heme oxygenase-1 silencing on the survival of acute myelocytic leukemia-M2 cells.

Wei S, Wang Y, Chai Q, Fang Q, Zhang Y, Wang J - Exp Ther Med (2015)

Bottom Line: The survival rate of the cells was significantly reduced by infection with pRNAi-siHO-1-GFP.HO-1 expression in the BMMNCs infected with pRNAi-siHO-1-GFP was downregulated, whereas caspase-3, -8 and -9 expression was upregulated compared with that in control BMMNCs.Kasumi-1 cells were successfully inoculated into nude mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, The Affiliated Hospital of Guiyang Medical College, Guiyang, Guizhou 550004, P.R. China ; Hematopoietic Stem Cell Transplantation Center of Guizhou Province, The Affiliated Hospital of Guiyang Medical College, Guiyang, Guizhou 550004, P.R. China ; Department of Clinical Biochemistry, The Affiliated Hospital of Guiyang Medical College, Guiyang, Guizhou 550004, P.R. China.

ABSTRACT

The aim of this study was to investigate the in vivo and in vitro effects of targeted heme oxygenase-1 (HO-1) silencing on the proliferation and apoptosis of human acute myelocytic leukemia (AML)-M2 cells. Bone marrow mononuclear cells (BMMNCs) were infected by pRNAi-siHO-1-GFP. The viability of the BMMNCs was determined by cell counting kit-8 (CCK-8) assay following daunorubicin (DNR) treatment. The apoptotic rate was detected by flow cytometry. The expression levels of HO-1 and apoptosis-related genes were detected by quantitative polymerase chain reaction (qPCR) and western blot analysis. An AML-M2 xenograft mouse model was established. The tumor formation outcomes and survival were observed. The leukocyte and platelet counts and hemoglobin levels were monitored, and the copy numbers of AML1/ETO fusion gene were detected by qPCR. pRNAi-siHO-1-GFP silenced the expression of HO-1. DNR inhibited cell viability in a time- and dose-dependent manner. The survival rate of the cells was significantly reduced by infection with pRNAi-siHO-1-GFP. HO-1 expression in the BMMNCs infected with pRNAi-siHO-1-GFP was downregulated, whereas caspase-3, -8 and -9 expression was upregulated compared with that in control BMMNCs. Kasumi-1 cells were successfully inoculated into nude mice. The rats inoculated with pRNAi-siHO-1-GFP-transfected Kasumi-1 cells succumbed to tumors more slowly and survived longer than those inoculated with untransfected Kasumi-1 cells. Furthermore, the leukocyte and platelet counts and hemoglobin levels were higher and the copy numbers of AML1/ETO fusion gene were lower in the former group. HO-1 gene silencing may promote the apoptosis of human M2 leukemic cells by inhibiting a caspase-dependent apoptotic pathway. Targeted silencing of HO-1 is able to inhibit the proliferation and infiltration of leukemic cells in nude mice and thus prolong their survival. The findings provide valuable experimental evidence for the molecular targeted therapy of M2 leukemia.

No MeSH data available.


Related in: MedlinePlus

Changes of body weight, leukocyte and platelet counts and hemoglobin levels, and Kaplan-Meier survival curve in the nude mouse model. (A) The Kasumi-1 cell-inoculated nude mouse model; left, Kasumi group; right, pRNAi-siHO-1-K group. (B) Volume of subcutaneous nodules in the nude mouse model; left, Kasumi group; right, pRNAi-siHO-1-K group. (C) Body weight changes of the blank, normal saline, Kasumi and pRNAi-siHO-1-Kasumi groups were recorded every 5 days. The body weights of the Kasumi group and the pRNAi-siHO-1-K group decreased from day 5, and that of the Kasumi group dropped to a greater extent. (D) Changes of leukocyte counts. The count of the Kasumi group reduced significantly (P<0.05). (E) Changes of hemoglobin levels. The hemoglobin level in the peripheral blood of the Kasumi group was significantly reduced (P<0.05). (F) Changes of platelet counts. The count of the Kasumi group was significantly lower than those of the other three groups (P<0.05). (G) Kaplan-Meier survival curves for evaluating the survival time of nude mice. The pRNAi-siHO-1-K group survived longer than the Kasumi group did. WBC, white blood cell; PLT, platelet; Hb, hemoglobin; HO-1, heme oxygenase-1.
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f7-etm-09-03-0931: Changes of body weight, leukocyte and platelet counts and hemoglobin levels, and Kaplan-Meier survival curve in the nude mouse model. (A) The Kasumi-1 cell-inoculated nude mouse model; left, Kasumi group; right, pRNAi-siHO-1-K group. (B) Volume of subcutaneous nodules in the nude mouse model; left, Kasumi group; right, pRNAi-siHO-1-K group. (C) Body weight changes of the blank, normal saline, Kasumi and pRNAi-siHO-1-Kasumi groups were recorded every 5 days. The body weights of the Kasumi group and the pRNAi-siHO-1-K group decreased from day 5, and that of the Kasumi group dropped to a greater extent. (D) Changes of leukocyte counts. The count of the Kasumi group reduced significantly (P<0.05). (E) Changes of hemoglobin levels. The hemoglobin level in the peripheral blood of the Kasumi group was significantly reduced (P<0.05). (F) Changes of platelet counts. The count of the Kasumi group was significantly lower than those of the other three groups (P<0.05). (G) Kaplan-Meier survival curves for evaluating the survival time of nude mice. The pRNAi-siHO-1-K group survived longer than the Kasumi group did. WBC, white blood cell; PLT, platelet; Hb, hemoglobin; HO-1, heme oxygenase-1.

Mentions: The tumor formation outcomes and nodule volume changes of the four nude mouse groups are presented in Fig. 7A and B. No tumors formed in the blank or normal saline groups. Tumors formed in the Kasumi group on day 6 after inoculation when the nodules were ~0.4 cm in diameter, and the nodules further grew to ~1.7 cm on day 15. The pRNAi-siHO-1-K group developed tumors on day 10 after inoculation, with ~0.3-cm-diameter nodules that grew to ~1.2 cm on day 19. The body weight changes of the four groups are shown in Fig. 7C. The body weights of the blank and normal saline groups were almost unchanged. By contrast, the Kasumi group underwent a sharp reduction in body weight. The pRNAi-siHO-1-K group underwent a similar body weight reduction to that of the Kasumi group over a longer time period.


In vivo and in vitro effects of heme oxygenase-1 silencing on the survival of acute myelocytic leukemia-M2 cells.

Wei S, Wang Y, Chai Q, Fang Q, Zhang Y, Wang J - Exp Ther Med (2015)

Changes of body weight, leukocyte and platelet counts and hemoglobin levels, and Kaplan-Meier survival curve in the nude mouse model. (A) The Kasumi-1 cell-inoculated nude mouse model; left, Kasumi group; right, pRNAi-siHO-1-K group. (B) Volume of subcutaneous nodules in the nude mouse model; left, Kasumi group; right, pRNAi-siHO-1-K group. (C) Body weight changes of the blank, normal saline, Kasumi and pRNAi-siHO-1-Kasumi groups were recorded every 5 days. The body weights of the Kasumi group and the pRNAi-siHO-1-K group decreased from day 5, and that of the Kasumi group dropped to a greater extent. (D) Changes of leukocyte counts. The count of the Kasumi group reduced significantly (P<0.05). (E) Changes of hemoglobin levels. The hemoglobin level in the peripheral blood of the Kasumi group was significantly reduced (P<0.05). (F) Changes of platelet counts. The count of the Kasumi group was significantly lower than those of the other three groups (P<0.05). (G) Kaplan-Meier survival curves for evaluating the survival time of nude mice. The pRNAi-siHO-1-K group survived longer than the Kasumi group did. WBC, white blood cell; PLT, platelet; Hb, hemoglobin; HO-1, heme oxygenase-1.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4316930&req=5

f7-etm-09-03-0931: Changes of body weight, leukocyte and platelet counts and hemoglobin levels, and Kaplan-Meier survival curve in the nude mouse model. (A) The Kasumi-1 cell-inoculated nude mouse model; left, Kasumi group; right, pRNAi-siHO-1-K group. (B) Volume of subcutaneous nodules in the nude mouse model; left, Kasumi group; right, pRNAi-siHO-1-K group. (C) Body weight changes of the blank, normal saline, Kasumi and pRNAi-siHO-1-Kasumi groups were recorded every 5 days. The body weights of the Kasumi group and the pRNAi-siHO-1-K group decreased from day 5, and that of the Kasumi group dropped to a greater extent. (D) Changes of leukocyte counts. The count of the Kasumi group reduced significantly (P<0.05). (E) Changes of hemoglobin levels. The hemoglobin level in the peripheral blood of the Kasumi group was significantly reduced (P<0.05). (F) Changes of platelet counts. The count of the Kasumi group was significantly lower than those of the other three groups (P<0.05). (G) Kaplan-Meier survival curves for evaluating the survival time of nude mice. The pRNAi-siHO-1-K group survived longer than the Kasumi group did. WBC, white blood cell; PLT, platelet; Hb, hemoglobin; HO-1, heme oxygenase-1.
Mentions: The tumor formation outcomes and nodule volume changes of the four nude mouse groups are presented in Fig. 7A and B. No tumors formed in the blank or normal saline groups. Tumors formed in the Kasumi group on day 6 after inoculation when the nodules were ~0.4 cm in diameter, and the nodules further grew to ~1.7 cm on day 15. The pRNAi-siHO-1-K group developed tumors on day 10 after inoculation, with ~0.3-cm-diameter nodules that grew to ~1.2 cm on day 19. The body weight changes of the four groups are shown in Fig. 7C. The body weights of the blank and normal saline groups were almost unchanged. By contrast, the Kasumi group underwent a sharp reduction in body weight. The pRNAi-siHO-1-K group underwent a similar body weight reduction to that of the Kasumi group over a longer time period.

Bottom Line: The survival rate of the cells was significantly reduced by infection with pRNAi-siHO-1-GFP.HO-1 expression in the BMMNCs infected with pRNAi-siHO-1-GFP was downregulated, whereas caspase-3, -8 and -9 expression was upregulated compared with that in control BMMNCs.Kasumi-1 cells were successfully inoculated into nude mice.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, The Affiliated Hospital of Guiyang Medical College, Guiyang, Guizhou 550004, P.R. China ; Hematopoietic Stem Cell Transplantation Center of Guizhou Province, The Affiliated Hospital of Guiyang Medical College, Guiyang, Guizhou 550004, P.R. China ; Department of Clinical Biochemistry, The Affiliated Hospital of Guiyang Medical College, Guiyang, Guizhou 550004, P.R. China.

ABSTRACT

The aim of this study was to investigate the in vivo and in vitro effects of targeted heme oxygenase-1 (HO-1) silencing on the proliferation and apoptosis of human acute myelocytic leukemia (AML)-M2 cells. Bone marrow mononuclear cells (BMMNCs) were infected by pRNAi-siHO-1-GFP. The viability of the BMMNCs was determined by cell counting kit-8 (CCK-8) assay following daunorubicin (DNR) treatment. The apoptotic rate was detected by flow cytometry. The expression levels of HO-1 and apoptosis-related genes were detected by quantitative polymerase chain reaction (qPCR) and western blot analysis. An AML-M2 xenograft mouse model was established. The tumor formation outcomes and survival were observed. The leukocyte and platelet counts and hemoglobin levels were monitored, and the copy numbers of AML1/ETO fusion gene were detected by qPCR. pRNAi-siHO-1-GFP silenced the expression of HO-1. DNR inhibited cell viability in a time- and dose-dependent manner. The survival rate of the cells was significantly reduced by infection with pRNAi-siHO-1-GFP. HO-1 expression in the BMMNCs infected with pRNAi-siHO-1-GFP was downregulated, whereas caspase-3, -8 and -9 expression was upregulated compared with that in control BMMNCs. Kasumi-1 cells were successfully inoculated into nude mice. The rats inoculated with pRNAi-siHO-1-GFP-transfected Kasumi-1 cells succumbed to tumors more slowly and survived longer than those inoculated with untransfected Kasumi-1 cells. Furthermore, the leukocyte and platelet counts and hemoglobin levels were higher and the copy numbers of AML1/ETO fusion gene were lower in the former group. HO-1 gene silencing may promote the apoptosis of human M2 leukemic cells by inhibiting a caspase-dependent apoptotic pathway. Targeted silencing of HO-1 is able to inhibit the proliferation and infiltration of leukemic cells in nude mice and thus prolong their survival. The findings provide valuable experimental evidence for the molecular targeted therapy of M2 leukemia.

No MeSH data available.


Related in: MedlinePlus