Danzhi Xiaoyaosan-containing Serum Reverses Energy Metabolic Reprogramming of MDA-MB-231 Breast Cancer Cells / 中国实验方剂学杂志
Chinese Journal of Experimental Traditional Medical Formulae
; (24): 8-14, 2022.
Article
en Zh
| WPRIM
| ID: wpr-940546
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WPRO
ABSTRACT
ObjectiveTo observe the effect of Danzhi Xiaoyaosan-containing serum on MDA-MB-231 breast cancer cells, and to find out whether the action mechanism is related to its intervention in energy metabolism. MethodThirty six-week-old male SD rats were randomly divided into the blank group, Danzhi Xiaoyaosan (8.99 g·kg-1) group, and Xihuangwan (0.55 g·kg-1) group. The serum was isolated after drug intervention for seven days. The cell viability was detected by methyl thiazolyl-tetrazolium (MTT) assay, the cell cycle by flow cytometry, and the apoptosis by Annenxin V/propidium iodide (PI) double staining. Following the determination of intracellular glucose content using the glucose testing kit, the expression of glucose transporter 1 (GLUT1) was measured by immunofluorescence staining. Seahorse XFe cell energy metabolism analyzer was used to detect the extracellular acidification rate (ECAR) and oxygen consumption rate (OCR). The expression levels of hexokinase 2 (HK2), pyruvate kinase M2 (PKM2), and lactate dehydrogenase A (LDHA) were assayed by Western blot. ResultCompared with the blank group, Danzhi Xiaoyaosan-containing serum inhibited the proliferation of MDA-MB-231 cells, with the best effect observed after intervention with 15% Danzhi Xiaoyaosan-containing serum for 48 h (P<0.01), blocked the MDA-MB-231 cells in G0/G1 phase(P<0.01), and down-regulated the GLUT1 expression, basal glycolysis, glycolysis capacity, glycolytic reserve, basal respiration, adenosine triphosphate (ATP) production, spare respiratory capacity(P<0.01), as well as the protein expression of HK2, PKM2, and LDHA in MDA-MB-231 cells(P<0.05,P<0.01). ConclusionDanzhi Xiaoyaosan-containing serum inhibits MDA-MB-231 cell proliferation, promotes apoptosis, and induces cell cycle arrest, which may be related to its reversal of energy metabolic reprogramming in MDA-MB-231 cells.
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WPRIM
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Zh
Revista:
Chinese Journal of Experimental Traditional Medical Formulae
Año:
2022
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Article