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Targeting cholesterol synthesis increases chemoimmuno-sensitivity in chronic lymphocytic leukemia cells.

Benakanakere I, Johnson T, Sleightholm R, Villeda V, Arya M, Bobba R, Freter C, Huang C - Exp Hematol Oncol (2014)

Bottom Line: We found that MEC-2 cells treated with cholesterol lowering agents (BIBB-515, YM-53601 or TAK-475) reduced 20% of total cellular cholesterol levels, but also significantly promoted CD-20 surface expression.More importantly, these cholesterol lowering agents also significantly enhanced chemoimmuno-sensitivity of the PBMCs from CLL patients.These results provide a novel strategy which could be applied to CLL treatment.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Hematology/Oncology, Department of Medicine, School of Medicine, University of Missouri, Columbia, MO 65212, USA.

ABSTRACT

Background: Cholesterol plays an important role in cancer development, drug resistance and chemoimmuno-sensitivity. Statins, cholesterol lowering drugs, can induce apoptosis, but also negatively interfere with CD-20 and rituximab-mediated activity. Our goal is to identify the alternative targets that could reduce cholesterol levels but do not interfere with CD-20 in chemo immunotherapy of chronic lymphocytic leukemia (CLL).

Methods: MEC-2 cells, a CLL cell line, and the peripheral blood mononuclear cells (PBMCs) from CLL patients were treated with cholesterol lowering agents, and analyzed the effect of these agents on cholesterol levels, CD-20 expression and distribution, and cell viability in the presence or absence of fludarabine, rituximab or their combinations.

Results: We found that MEC-2 cells treated with cholesterol lowering agents (BIBB-515, YM-53601 or TAK-475) reduced 20% of total cellular cholesterol levels, but also significantly promoted CD-20 surface expression. Furthermore, treatment of cells with fludarabine, rituximab or their combinations in the presence of BIBB-515, YM-53601 or TAK-475 enhanced MEC-2 cell chemoimmuno-sensitivity measured by cell viability. More importantly, these cholesterol lowering agents also significantly enhanced chemoimmuno-sensitivity of the PBMCs from CLL patients.

Conclusion: Our data demonstrate that BIBB-515, YM53601 and TAK-475 render chemoimmuno-therapy resistant MEC-2 cells sensitive to chemoimmuno-therapy and enhance CLL cell chemoimmuno-sensitivity without CD-20 epitope presentation or its downstream signaling. These results provide a novel strategy which could be applied to CLL treatment.

No MeSH data available.


Related in: MedlinePlus

Effect of TAK-475 on CD-20 expression and chemoimmuno-sensitivity. MEC-2 cells were treated with 10 μM TAK-475 (T) for 3 days, processed for double immunostaining, and photographed by confocal microscope (A-D). The results represent three experiments performed with duplicate samples. The cells were also treated with 10 μM fludarabine (F), 10 μg/ml rituximab (R) or their combinations (F + R) in the presence or absence of 10 μM TAK-475 for 3 days, and then analyzed cell viability by MTT assay (E). The values of treated with TAK-475 samples (n = 16) were statistically different from the controls (C). *P < 0.05. **P < 0.01.
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Figure 6: Effect of TAK-475 on CD-20 expression and chemoimmuno-sensitivity. MEC-2 cells were treated with 10 μM TAK-475 (T) for 3 days, processed for double immunostaining, and photographed by confocal microscope (A-D). The results represent three experiments performed with duplicate samples. The cells were also treated with 10 μM fludarabine (F), 10 μg/ml rituximab (R) or their combinations (F + R) in the presence or absence of 10 μM TAK-475 for 3 days, and then analyzed cell viability by MTT assay (E). The values of treated with TAK-475 samples (n = 16) were statistically different from the controls (C). *P < 0.05. **P < 0.01.

Mentions: TAK-475 (Lapaquistat) is another squalene synthase inhibitor that has been used as a cholesterol lowering drug in Phase III clinical trials in Europe and the United States[39]. To investigate the effect of TAK-475 on chemoimmuno-sensitivity, we firstly treated MEC-2 cells with 10 μM TAK-475 for 3 days, and explored its effect on CD-20 expression and distribution. As shown in Figure 6, both CD-20 staining and membrane association are significantly increase in TAK-475 treated (Figure 6C and D) compared with those controls (Figure 6A and B). Next, we analyze the effect of TAK-475 on MEC-2 cell chemoimmuno-sensitivity. As shown in Figure 6E, TAK-475 enhances chemoimmuno-sensitivity of fludarabine, rituximab and their combinations. With TAK-475, cell viability was decreased from 72% to 40% with fludarabine treatment alone, from 80% to 59% with rituximab treatment alone, and from 41% to 26% with their combination treatment.


Targeting cholesterol synthesis increases chemoimmuno-sensitivity in chronic lymphocytic leukemia cells.

Benakanakere I, Johnson T, Sleightholm R, Villeda V, Arya M, Bobba R, Freter C, Huang C - Exp Hematol Oncol (2014)

Effect of TAK-475 on CD-20 expression and chemoimmuno-sensitivity. MEC-2 cells were treated with 10 μM TAK-475 (T) for 3 days, processed for double immunostaining, and photographed by confocal microscope (A-D). The results represent three experiments performed with duplicate samples. The cells were also treated with 10 μM fludarabine (F), 10 μg/ml rituximab (R) or their combinations (F + R) in the presence or absence of 10 μM TAK-475 for 3 days, and then analyzed cell viability by MTT assay (E). The values of treated with TAK-475 samples (n = 16) were statistically different from the controls (C). *P < 0.05. **P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4231203&req=5

Figure 6: Effect of TAK-475 on CD-20 expression and chemoimmuno-sensitivity. MEC-2 cells were treated with 10 μM TAK-475 (T) for 3 days, processed for double immunostaining, and photographed by confocal microscope (A-D). The results represent three experiments performed with duplicate samples. The cells were also treated with 10 μM fludarabine (F), 10 μg/ml rituximab (R) or their combinations (F + R) in the presence or absence of 10 μM TAK-475 for 3 days, and then analyzed cell viability by MTT assay (E). The values of treated with TAK-475 samples (n = 16) were statistically different from the controls (C). *P < 0.05. **P < 0.01.
Mentions: TAK-475 (Lapaquistat) is another squalene synthase inhibitor that has been used as a cholesterol lowering drug in Phase III clinical trials in Europe and the United States[39]. To investigate the effect of TAK-475 on chemoimmuno-sensitivity, we firstly treated MEC-2 cells with 10 μM TAK-475 for 3 days, and explored its effect on CD-20 expression and distribution. As shown in Figure 6, both CD-20 staining and membrane association are significantly increase in TAK-475 treated (Figure 6C and D) compared with those controls (Figure 6A and B). Next, we analyze the effect of TAK-475 on MEC-2 cell chemoimmuno-sensitivity. As shown in Figure 6E, TAK-475 enhances chemoimmuno-sensitivity of fludarabine, rituximab and their combinations. With TAK-475, cell viability was decreased from 72% to 40% with fludarabine treatment alone, from 80% to 59% with rituximab treatment alone, and from 41% to 26% with their combination treatment.

Bottom Line: We found that MEC-2 cells treated with cholesterol lowering agents (BIBB-515, YM-53601 or TAK-475) reduced 20% of total cellular cholesterol levels, but also significantly promoted CD-20 surface expression.More importantly, these cholesterol lowering agents also significantly enhanced chemoimmuno-sensitivity of the PBMCs from CLL patients.These results provide a novel strategy which could be applied to CLL treatment.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Hematology/Oncology, Department of Medicine, School of Medicine, University of Missouri, Columbia, MO 65212, USA.

ABSTRACT

Background: Cholesterol plays an important role in cancer development, drug resistance and chemoimmuno-sensitivity. Statins, cholesterol lowering drugs, can induce apoptosis, but also negatively interfere with CD-20 and rituximab-mediated activity. Our goal is to identify the alternative targets that could reduce cholesterol levels but do not interfere with CD-20 in chemo immunotherapy of chronic lymphocytic leukemia (CLL).

Methods: MEC-2 cells, a CLL cell line, and the peripheral blood mononuclear cells (PBMCs) from CLL patients were treated with cholesterol lowering agents, and analyzed the effect of these agents on cholesterol levels, CD-20 expression and distribution, and cell viability in the presence or absence of fludarabine, rituximab or their combinations.

Results: We found that MEC-2 cells treated with cholesterol lowering agents (BIBB-515, YM-53601 or TAK-475) reduced 20% of total cellular cholesterol levels, but also significantly promoted CD-20 surface expression. Furthermore, treatment of cells with fludarabine, rituximab or their combinations in the presence of BIBB-515, YM-53601 or TAK-475 enhanced MEC-2 cell chemoimmuno-sensitivity measured by cell viability. More importantly, these cholesterol lowering agents also significantly enhanced chemoimmuno-sensitivity of the PBMCs from CLL patients.

Conclusion: Our data demonstrate that BIBB-515, YM53601 and TAK-475 render chemoimmuno-therapy resistant MEC-2 cells sensitive to chemoimmuno-therapy and enhance CLL cell chemoimmuno-sensitivity without CD-20 epitope presentation or its downstream signaling. These results provide a novel strategy which could be applied to CLL treatment.

No MeSH data available.


Related in: MedlinePlus