Preliminary Study on the Anti-Proliferative Mechanism of Hesperetin against MDA-MB-231 Cells Based on RNA-Seq Analysis

ORIGINAL ARTICLE
Yu-Zhen Ma, Shuang-Shuang Sun, Guang-Zhou Zhou

 
For citation: Ma Y-Z, Sun S-S, Zhou G-Z. Preliminary Study on the Anti-Proliferative Mechanism of Hesperetin against MDA-MB-231 Cells Based on RNA-Seq Analysis. International Journal of Biomedicine. 2026;16(2):157-162. doi:10.21103/Article16(2)_OA1
 
Originally published June 5, 2026

Abstract: 

Background: Hesperetin is a natural dihydroflavonoid compound with multiple pharmacological activities. Previous studies have shown that hesperetin targets multiple cellular proteins, such as cell cycle regulation, apoptosis, metastasis, tyrosine kinases, growth factor receptors, estrogen metabolism, and antioxidant-related proteins, to inhibit tumor formation.
Methods and Results: In this study, we treated the triple-negative breast cancer cell line MDA-MB-231 with hesperetin for 24 h and extracted total RNA for RNA-seq analysis, which revealed that 1203 genes were up-regulated and 1997 genes were down-regulated. GO clustering and KEGG enrichment analysis indicated that most of these genes played important roles in tumorigenesis and various cellular signaling pathways. Four genes related to tumor cell apoptosis and migration, including NR4A1, CSF2, KDR, and LRRK2, were selected for real-time RT-PCR analysis for validation. The mRNA level changes were consistent with the transcriptome sequencing results. Further western blot analysis of changes in the expression of the orphan nuclear receptor NR4A1 and the apoptosis-related protein Bcl-2 showed that they may be involved in hesperetin-induced apoptosis in MDA-MB-231 cells. The current research results provide a preliminary basis for further elucidating the molecular mechanism of hesperetin against breast cancer and for developing future clinical drugs based on hesperetin.

Keywords: 
breast cancer • hesperetin • transcriptome sequencing • orphan nuclear receptor protein • molecular mechanism
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Received February 14, 2026.
Accepted March 31, 2026.
© 2026 The Author(s). International Journal of Biomedicine is published by IMRDC.
This is an open access article under the CC BY-NC-ND 4.0 license.