Antioxidant and Anti-tumor Effect and Mechanisms of Andrographis paniculata Polysaccharide on Human Cervical Cancer HeLa Cells
10.13422/j.cnki.syfjx.20240821
- VernacularTitle:苦胆草多糖抗氧化及抗人宫颈癌HeLa细胞的作用及机制
- Author:
Lijin HUANG
1
;
Zilin LI
1
;
Ziyan YANG
1
;
Han WANG
1
;
Guiyuan CHEN
1
Author Information
1. Basic Medicine School， Dali University， Dali 671000，China
- Publication Type:Journal Article
- Keywords:
Andrographis paniculata polysaccharide;
cervical cancer HeLa cells;
anti-tumor;
antioxidant;
apoptosis
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2024;30(18):80-88
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the antioxidant and anti-human cervical cancer HeLa cell effect and mechanisms of Andrographis paniculata polysaccharide （APP）. MethodCell function assays were conducted to assess the effects of APP （400， 450， 500 mg·L-1） on the proliferation， migration， and invasion of HeLa cells using the cell counting kit-8 （CCK-8） assay， scratch assay， and Transwell assay. Molecular mechanism experiments were conducted to detect the effects of APP on HeLa cell apoptosis and cell cycle-related mRNA and protein expression using flow cytometry， real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR）， and Western blot analysis. The antioxidant activity of APP was tested using DPPH+， OH-， and reducing power assays. ResultCompared with the blank group， APP （400， 450， 500 mg·L-1） significantly inhibited the migration， proliferation， and invasion abilities of HeLa cells in a time- and dose-dependent manner （P<0.05， P<0.01）. Flow cytometry with propidium iodide （PI） single staining was used to detect cell cycle. The results showed that compared with the blank group， the proportion of HeLa cells in the G2/M phase increased after 48 hours of treatment with APP （400， 450， 500 mg·L-1）， indicating that APP can arrest HeLa cells in the G2/M phase. Flow cytometry with fluorescein isothiocyanate （Annexin V-FITC）/PI apoptosis kit was used to detect cell apoptosis. Compared with the blank group， the proportion of early and late apoptotic HeLa cells increased in a dose-dependent manner after 48 hours of APP （400， 450， 500 mg·L-1） treatment （P<0.05， P<0.01）， indicating that APP promotes HeLa cell apoptosis. The results of Real-time PCR and Western blot assay showed that compared with the blank group， after 48 hours of APP （400， 450， 500 mg·L-1） treatment resulted in decreased mRNA and protein expression of cell cycle-dependent kinase-1 （CDK-1）， Cyclin B1， and B-cell lymphoma-2 （Bcl-2）， and increased mRNA and protein expression of cysteine aspartate protease （Caspase）-3， Caspase-8， Caspase-9， and Bcl-2-associated X protein （Bax） （P<0.05， P<0.01）. These findings were consistent with the flow cytometry results and showed a dose-dependent effect. In vitro antioxidant tests demonstrated that different concentrations of APP （50-1 000 mg·L-1） were able to scavenge DPPH+ and OH- radicals， indicating certain antioxidant activity. ConclusionAPP possesses antioxidant activity and can inhibit the viability of HeLa cells while promoting their apoptosis.
