Effect of Pulsatilla Saponin A on Proliferation and Apoptosis of Burkitt Lymphoma Cells Based on JAK2/STAT3 Signaling Pathway
10.13422/j.cnki.syfjx.20221225
- VernacularTitle:基于JAK2/STAT3信号通路探讨白头翁皂苷A对Burkitt淋巴瘤细胞增殖和凋亡的影响
- Author:
Ning LIU
1
;
Jie BAI
1
;
Hui YU
2
;
Xiaoli CHEN
2
;
Xiangtu KONG
2
;
Haiwen NI
2
Author Information
1. The First School of Clinical Medicine of Nanjing University of Chinese Medicine ，Nanjing 210023， China
2. Jiangsu Province Hospital of Chinese Medicine（Affiliated Hospital of Nanjing University of Chinese Medicine）， Nanjing 210029， China
- Publication Type:Journal Article
- Keywords:
pulsatilla saponin A;
Burkitt lymphoma;
Raji cells;
proliferation;
apoptosis;
janus kinase 2/signal transducer and activator of transcription 3 （JAK2/STAT3） signaling pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(20):71-77
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effect of pulsatilla saponin A （PSA） on proliferation and apoptosis of human Burkitt lymphoma （BL） cell line Raji cells and expression of related pathway proteins. MethodWith Raji cells as the research object， the cell proliferation was detected by cell counting kit-8 （CCK-8） method， and the half-maximal inhibitory concentration （IC50） values of 24 h， 48 h and 72 h were calculated to be 19.77， 18.31， 16.70 μmol·L-1， respectively. In subsequent related experiments， 0， 8， 16， 32 μmol·L-1 PSA were selected according to the IC50 value of Raji cells treated with PAS for 72 h. After 0， 8， 16， 32 μmol·L-1 PSA acted on Raji cells for 24， 48， 72 h， the optical density values of cell growth curve were detected by CCK-8 method. The zymogen activities of cysteine aspartate-specific protease （Caspase）-3， Caspase-8 and Caspase-9 in Raji cells treated with 0， 8， 16 and 32 μmol·L-1 PSA for 24 h were measured by Caspase-3， Caspase-8 and Caspase-9 colorimetric assay kit. The apoptosis rate and cell cycle of Raji cells treated with different concentrations of PSA after 24 h were detected by flow cytometry. The expression of B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， cleaved poly（ADP-ribose） polymerase （cleaved PARP）， cleaved cysteinyl aspartate-specific protease-3 （cleaved Caspase-3） apoptosis related protein and Janus kinase 2 （JAK2）， signal transducer and activator of transcription 3 （STAT3）， phosphorylated-JAK2 （p-JAK2）， and phosphorylated- STAT3 （p-STAT3） pathway proteins in Raji cells after 24 h of treatment with 0， 8， 16 and 32 μmol·L-1 PSA were tested by Western blot. ResultCompared with control group， decreased cell survival rate， inhibited cell proliferation， activated zymogens of Caspase-3， Caspase-8 and Caspase-9 （P<0.01）， increased apoptosis （P<0.05， P<0.01）， and enhanced cell cycle arrest in Gap phase 2 （G2） were observed in 8， 16 and 32 μmol·L-1 PSA groups（P<0.05， P<0.01）. Compared with control group， cells treated with 8， 16 and 32 μmol·L-1 PSA had lower expression of Bcl-2， p-JAK2， p-STAT3 proteins （P<0.05， P<0.01）， and higher expression of Bax， cleaved PARP and cleaved Caspase-3 protein （P<0.01）， while no significant change was found in the expression of JAK2 and STAT3 proteins. ConclusionPSA could inhibit proliferation and induce apoptosis of Raji cells， and its potential mechanism might be related to the regulation of JAK2/STAT3 signaling pathway.
