Mechanism analysis of the apoptosis in chronic myeloid leukemia K562 cells induced by total alkaloids of Gelsemium elegans

WANG Wenyi; TAN Xinghui; LU Yi; LI Desen; WU Shuisheng

doi:10.13506/j.cnki.jpr.2024.02.001

Abstract：Objective To investigate the inhibitory influence of total alkaloids of Gelsemium elegans(TAG)on chronic myeloid leukemia K562 cells and its mechanism,so as to provide experimental basis for the use of GE in the treatment of CML.Methods K562 cells were treated with different concentrations of TAG(25,50,100,200,400 μg·mL-1),and the effect of TAG on K562 cell viability was determined by MTT assay.The morphology of K562 cells was observed by inverted phase contrast microscope.DAPI staining was applied to observe the nuclear morphological changes of K562.Annexin V FITC/PI flow cytometry was used to detect the apoptosis of K562 cells.Caspase-3 activity was measured by colorimetry.RT-PCR was applied to detect the expression levels of Bax and Bcl-2 genes.Results The inhibitory effect of TAG on K562 cells was time and dose dependent,and the IC50 was 122 μg·mL-1 at 24 h;After the TAG intervened K562 cells,the cell morphology was gradually irregular,the cytoplasm was unclear,the nucleus was shrunk,and finally the integrity of the cell membrane was destroyed;DAPI staining showed that the cell volume was reduced and shrunk,the nucleus was pyknotic,and typical apoptotic bodies were formed;The results of Annexin V FITC/PI flow cytometry showed that the apoptosis of K562 cells was mainly early withering,and there was a dose effect relationship;The activity of caspase-3 in K562 cells was in-creased after 24 h treatment with disparate concentrations of TAG;RT-PCR testing report exhibit that expression of Bcl-2 was down regulated and Bax expression was up regulated,both of which showed a dose effect relationship.Conclusion The TAG can up regulate the expression of Bax gene,down regulate the expression of Bcl-2 gene,activate caspase-3,thereby inducing the early apoptosis of K562 cells and inhibiting their cell viability.