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Effect and Mechanism of Xianlian Jiedu Prescription Against Proliferation of Colorectal Cancer Cells in Hypoxic Microenvironment / 中国实验方剂学杂志
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 79-85, 2022.
Article in Chinese | WPRIM | ID: wpr-940389
ABSTRACT
ObjectiveTo study the effect of Xianlian Jiedu prescription (XLJDP) on the activation of nuclear transcription factor-κB (NF-κB) signaling pathway induced by bromodomain-containing protein 4 (Brd4) in hypoxic microenvironment and to explore its mechanism in inhibiting the proliferation of colorectal cancer HT-29 cells. MethodThe human colorectal cancer HT-29 cells were cultured in a hypoxic incubator or normoxia incubator and treated with XLJDP at 0.8,1,1.2,1.6,3.2,6.4,and 12.8 g·L-1 for 48 h, respectively. Following the detection of cell vitality using methyl thiazolyl tetrazolium (MTT) colorimetry, the effects of XLJDP (1.25,2.5,and 5 g·L-1) on the cell mitochondrial membrane potential were determined using a fluorescent probe (JC-1), and the apoptosis of colorectal cancer HT-29 cells was detected by flow cytometry. The cell colony formation assay and 5-ethynyl-2'-deoxyuridine (EDU) staining were conducted to test the proliferation of colorectal cancer HT-29 cells. The Western blot was carried out to measure the expression levels of Brd4 and its downstream relevant proteins such as c-Myc and hexamethylene bisacetamide-inducible protein 1 (HEXIM1), as well as the effects of XLJDP on related proteins in the NF-κB signaling pathway. ResultCompared with the blank control group, XLJDP at 0.8,1,1.2,1.6,3.2,6.4,and 12.8 g·L-1 inhibited the vitality of colorectal cancer HT-29 cells (P<0.05 , P<0.01), with the median inhibitory concentration (IC50) under the hypoxic condition higher than that under the normoxia condition. Compared with the blank control group, XLJDP at 1.25,2.5,and 5 g·L-1 significantly decreased the mitochondria membrane potential, enhanced the apoptosis (P<0.05,P<0.01), and lowered the number of cell colonies and also the EDU-positive cells (P<0.05, P<0.01). The results of Western blot showed that compared with the blank control group, XLJDP at 1.25,2.5,and 5 g·L-1 down-regulated Brd4, c-Myc, p-NF-κB p65, and p-IκBα protein expression to varying degrees and up-regulated the expression of HEXIM1 (P<0.05, P<0.01). ConclusionIn the hypoxic microenvironment, XLJDP inhibits the proliferation of colorectal cancer HT-29 cells regulated by Brd4, which may be related to its inhibition of the activation of NF-κB signaling pathway.

Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2022 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2022 Type: Article