Xiaoli Wang1, Ruidong Liu1, Yanxia Wang2, Hengjuan Cai3, Lei Zhang4. 1. Department of Clinical Laboratory, People's Hospital of Laiwu Laiwu, China. 2. Department of Cardiology, People's Hospital of Zhangqiu Jinan, China. 3. Department of Neurology, People's Hospital of Zhangqiu Jinan, China. 4. Department of Clinical Laboratory, Qingdao Women and Children's Hospital Qingdao, China.
Abstract
AIMS AND BACKGROUND: Up-regulation of clusterin is associated with the survival and progression of various malignancies, and down-regulation of clusterin promotes apoptosis and inhibits invasion. The aim of this study was to explore the effect of clusterin small interference RNA (siRNA) on the proliferation, apoptosis and invasion of HL-60 acute myeloid leukemia (AML) cells. METHODS: siRNA transfection was performed using Lipofectamine™2000 reagent. Relative protein expressions were quantified by Western blot. Trypan blue assay was performed to assess tumor cell proliferation after siRNA transfection. The cytotoxic effect of clusterin siRNA on leukemic cells was measured using MTT assay. Apoptosis was detected using fluorescence microscopy assay. Migration and invasion was detected after clusterin was silenced. RESULTS: Clusterin siRNA clearly lowered clusterin protein levels in a time- dependent manner, leading to marked inhibition of cell survival, proliferation and invasion. Furthermore, clusterin down-regulation significantly enhanced the extent of HL-60 apoptotic cells. CONCLUSIONS: Our results suggest that the down-regulation of clusterin by siRNA can effectively trigger apoptosis and inhibit the proliferation and invasion of leukemic cells. Therefore, clusterin siRNA may be a potent adjuvant in AML therapy.
AIMS AND BACKGROUND: Up-regulation of clusterin is associated with the survival and progression of various malignancies, and down-regulation of clusterin promotes apoptosis and inhibits invasion. The aim of this study was to explore the effect of clusterin small interference RNA (siRNA) on the proliferation, apoptosis and invasion of HL-60 acute myeloid leukemia (AML) cells. METHODS: siRNA transfection was performed using Lipofectamine™2000 reagent. Relative protein expressions were quantified by Western blot. Trypan blue assay was performed to assess tumor cell proliferation after siRNA transfection. The cytotoxic effect of clusterin siRNA on leukemic cells was measured using MTT assay. Apoptosis was detected using fluorescence microscopy assay. Migration and invasion was detected after clusterin was silenced. RESULTS:Clusterin siRNA clearly lowered clusterin protein levels in a time- dependent manner, leading to marked inhibition of cell survival, proliferation and invasion. Furthermore, clusterin down-regulation significantly enhanced the extent of HL-60 apoptotic cells. CONCLUSIONS: Our results suggest that the down-regulation of clusterin by siRNA can effectively trigger apoptosis and inhibit the proliferation and invasion of leukemic cells. Therefore, clusterin siRNA may be a potent adjuvant in AML therapy.
Entities:
Keywords:
Acute myeloid leukemia; apoptosis; clusterin; invasion; small interference RNA
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