Effect of Stemona tuberosa Alkaloids on Apoptosis of SMMC-7721 Cells and Expression of Bcl-2， Bax， and Cleaved Caspase-3
10.13422/j.cnki.syfjx.20211921
- VernacularTitle:对叶百部总生物碱对人肝癌SMMC-7721细胞凋亡及Bcl-2，Bax和cleaved Caspase-3蛋白表达的影响
- Author:
Si LIN
1
;
Hua ZHU
1
;
Hui-zhen QIN
1
;
Xiao-xun WANG
1
;
Li-ba XU
2
;
Ling-yu DENG
1
Author Information
1. Guangxi University of Chinese Medicine，Nanning 530200，China
2. Guangxi Key Laboratory of Zhuang and Yao Ethnic Medicine，Guangxi University of Chinese Medicine，Nanning 530200，China
- Publication Type:Research Article
- Keywords:
Stemona tuberosa alkaloids;
human hepatoma;
SMMC-7721 cells;
apoptosis;
B lymphocytoma-2 （Bcl-2）;
Bcl-2-associated X protein （Bax）;
cleaved cysteinyl aspartate-specific protease-3 （cleaved Caspase-3）
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(19):73-79
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of Stemona tuberosa alkaloids on the apoptosis of human hepatoma SMMC-7721 cells and the expression of apoptosis-related proteins including B lymphocytoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and cleaved cysteinyl aspartate-specific protease-3 （cleaved Caspase-3）. Method:SMMC-7721 cells were routinely cultured， passaged， and treated with various concentrations （50， 75， 112， 167， and 250 mg·L-1） of S. tuberosa alkaloids， while those in the blank control group were only treated with 10% fetal bovine serum. The cell proliferation was determined by tetrazolium bromide （MTT） colorimetry and colony assay and the cell apoptosis by Hoechst 33258 staining. The protein expression levels of Bcl-2， Bax， and cleaved Caspase-3 were detected by Western blot. Result:S. tuberosa alkaloids inhibited the proliferation of SMMC-7721 cells， and the inhibition rate was significantly increased in comparison with that in the blank control group （P<0.01）， with the half maximal inhibitory concentrations （IC50） at 24 h， 48 h， and 72 h being （173.36±8.75）， （112.14±16.50）， and （96.41±2.60）mg·L-1， respectively. The cell colony-inhibitory activity was significantly increased in a dose-dependent manner （P<0.01）. Compared with the blank control group， S. tuberosa alkaloids promoted the apoptosis of SMMC-7721 cells， manifested as increased number of apoptotic cells and elevated apoptotic rate （P<0.01）. The typical morphological changes such as brightly blue-fluorescent condensed nuclei， cytoplasmic shrinking， and karyopyknosis were found under the upright fluorescence microscope. As revealed by comparison with the blank control group， the expression of Bcl-2 was significantly down-regulated （P<0.01）， while the protein expression levels of pro-apoptotic protein Bax and cleaved Caspase-3 in the 75， 112， 167， and 250 mg·L-1 S. tuberosa alkaloids groups were significantly up-regulated （P<0.01）. Conclusion:S. tuberosa alkaloids inhibit the proliferation of SMMC-7721 cells and promote their apoptosis possibly by inhibiting Bcl-2 protein expression and promoting Bax and cleaved Caspase-3 protein expression.
