Effects of Honokiol on Proliferation and Apoptosis on U937 Cells
Ling Pan
Abstract
Ling Pan
Abstract
【Objective】 To investigate the anti-proliferative and apoptosis effect induced by Honokiol(HNK) on human myeloid leukemia cell line U937 cells in vitro.【Methods】 After treated with different concentration of HNK,Hoechst33342 fluorescent staining was used to detect cell apoptosis;the growth inhibition ration of U937 cells and PBMCs were analyzed by MTT assay;the apoptosis ration was detected by flow cytometry;mitochondrial membrane potential was explored by rhodamine 123 stain;Caspase3/7 protein activity kit was used to test the Caspase3/7 activity;the Caspase-3 and Caspase-7 mRNA levels were detected by real-time fluorescent relative-quantification reverse transcriptional PCR(FQ-PCR).【Results】 Honokiol could significantly inhibit the proliferation of U937 cells in terms of the indexes of IC50/U937 11.8 μg/mL and IC50/PBMCs 40.3 μg/mL,and the anti-proliferative effect was in a time and concentration dependent manner;Flow cytometry analysis manifested that Honokiol could induce U937cells apoptosis by Annexin V/PI double Annexin V/PI fluorescein stain;Honokiol significantly inhibited the mitochondrial membrane potential of U937 cells and enhanced the ability of Caspase3/7 and the mRNA expression levels,but not the PBMCs.【Conclusion】 HNK can inhibit U937 cells proliferation and induce cells apoptosis via activating Caspase 3/7.
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【Objective】 To investigate the anti-proliferative and apoptosis effect induced by Honokiol(HNK) on human myeloid leukemia cell line U937 cells in vitro.【Methods】 After treated with different concentration of HNK,Hoechst33342 fluorescent staining was used to detect cell apoptosis;the growth inhibition ration of U937 cells and PBMCs were analyzed by MTT assay;the apoptosis ration was detected by flow cytometry;mitochondrial membrane potential was explored by rhodamine 123 stain;Caspase3/7 protein activity kit was used to test the Caspase3/7 activity;the Caspase-3 and Caspase-7 mRNA levels were detected by real-time fluorescent relative-quantification reverse transcriptional PCR(FQ-PCR).【Results】 Honokiol could significantly inhibit the proliferation of U937 cells in terms of the indexes of IC50/U937 11.8 μg/mL and IC50/PBMCs 40.3 μg/mL,and the anti-proliferative effect was in a time and concentration dependent manner;Flow cytometry analysis manifested that Honokiol could induce U937cells apoptosis by Annexin V/PI double Annexin V/PI fluorescein stain;Honokiol significantly inhibited the mitochondrial membrane potential of U937 cells and enhanced the ability of Caspase3/7 and the mRNA expression levels,but not the PBMCs.【Conclusion】 HNK can inhibit U937 cells proliferation and induce cells apoptosis via activating Caspase 3/7.
Key concepts: Flow cytometry, Apoptosis, Honokiol, Annexin, U937 cell, Molecular biology, Cell growth, Chemistry