|Table of Contents|

Inhibition of triple-negative breast cancer cell proliferation by synergistic induction of nucleolar stress by Flavokawain B and Actinomycin D

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2024 23
Page:
4432-4442
Research Field:
Publishing date:

Info

Title:
Inhibition of triple-negative breast cancer cell proliferation by synergistic induction of nucleolar stress by Flavokawain B and Actinomycin D
Author(s):
Linghui; GAO Qixue; HE Yesong; LI Xuesen
Institute for Cancer Medicine,School of Basic Medical Sciences,Southwest Medical University,Sichuan Luzhou 646000,China.
Keywords:
triple-negative breast cancer; Flavokawain B; Actinomycin D; nucleolar stress; cell apoptosis
PACS:
R737.9
DOI:
10.3969/j.issn.1672-4992.2024.23.003
Abstract:
Objective:To investigate the synergistic inhibition of triple-negative breast cancer cell proliferation by Flavokawain B(FKB) and Actinomycin D(ActD),as well as their synergistic anticancer mechanism.Methods:Cultivate SUM159PT and HS578T triple-negative breast cancer cells,and employ the MTT assay to assess the inhibitory effects of different concentrations of FKB and ActD monotherapies,as well as appropriate concentrations of combination therapy,on the proliferation of these two types of cancer cells.To utilize flow cytometry to examine the effects of FKB and ActD monotherapies,as well as combination therapy,on cell cycle progression and apoptosis in the two types of triple-negative breast cancer cells.Conduct RT-qPCR experiments to analyze the changes in free rRNA that were not assembed into ribosomes following treatment with FKB and ActD monotherapies,as well as combination therapy,in the two types of triple-negative breast cancer cells.Perform cell immunofluorescence experiments to observe the nuclear localization of NPM1 protein in the two types of triple-negative breast cancer cells after treatment with FKB and ActD monotherapies,as well as combination therapy,in order to identify cellular nucleolar stress induction.Finally,to utilize Western blot experiments to assess changes in cell cycle and apoptosis-related proteins in the two types of triple-negative breast cancer cells following treatment with FKB and ActD monotherapies,as well as combination therapy.Results:FKB and ActD synergistically inhibited cell proliferation in two types of triple-negative breast cancer cells,and flow cytometry results showed that the combination therapy significantly induced more apoptosis in triple-negative breast cancer cells(P＜0.01) and affected cell cycle progression,and that the combination’s inhibition of cell proliferation was attenuated by inhibition of apoptotic signaling pathways using apoptosis inhibitors and by controlling the cell cycle using serum deprivation(P＜0.01).RT-qPCR results indicated that combination therapy led to a significant accumulation of both mature and immature 18S and 28S rRNA in triple-negative breast cancer cells (P＜0.01),suggesting severe disruption of ribosomal RNA splicing or assembly.Cell immunofluorescence results showed that NPM1 protein in triple-negative breast cancer cells exhibited a diffuse distribution throughout the nucleus after combination therapy,indicating that the disruption of ribosomal RNA splicing or assembly triggered a nucleolar stress response in triple-negative breast cancer cells.Western blot results demonstrated that combination therapy significantly upregulated p21 protein,downregulated Bcl2 protein,and induced cleavage of Caspase3 and PARP proteins.Conclusion:FKB and ActD may synergistically disrupt ribosomal RNA splicing or assembly,destabilizing nucleolar homeostasis and triggering a nucleolar stress response.This,in turn,inducing upregulation of downstream p21 protein,leading to apoptosis of triple-negative breast cancer cells and triggering cell cycle arrest,thereby exerting a synergistic inhibitory effect on the proliferation of triple-negative breast cancer cells.

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Last Update: 2024-11-01