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ELF3 promotes gemcitabine resistance through PKMYT1/CDK1 signaling pathway in gallbladder cancer
Linhua Yang1,2,3,4, Hui Wang1,2,3,4, Miaomiao Guo5
1Department of Biliary-Pancreatic Surgery, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China.
Background:
Gemcitabine is the standard treatment for gallbladder cancer (GBC) patients, and the development of resistance frequently limits its efficacy. However, the molecular features and mechanisms of gemcitabine resistance (Gem-R) in GBC cells remain unknown. Herein, we aimed to explore the role of ELF3 in Gem-R of GBC, including the underlying mechanisms.
Methods:
RNA sequencing was used to screen the essential genes related to the generation of Gem-R in GBC tissues. The correlation between Gem-R and ELF3 expression was identified in GDSC, GEO database, GBC tissues, and 3 GBC cell lines. Immunohistochemical staining, quantitative real-time polymerase chain reaction, and western blot were used to examine the expression of ELF3, PKMYT1, and CDK1. Luciferase reporter assays were used to identify the binding site of ELF3 in the PKMYT1 promoter region. CCK-8 assay and clonogenic survival assays were used to evaluate the sensitivity of gemcitabine in GBC cells. A GBC xenograft model was used to evaluate the influence of ELF3 on the therapeutic effect of gemcitabine.
Results:
A consistently positive correlation between ELF3 expression and Gem-R, both in newly generated GBC RNA-seq data and in the datasets from GDSC and GEO. Gem-R in GBC cells was facilitated by ELF3 overexpression, whereas ELF3 knockdown had the opposite effect. In vivo experiments further proved that reducing ELF3 expression promoted the gemcitabine sensitivity of GBC cells and extended the survival time of mice that received orthotopic xenografted tumors. Mechanistically, ELF3 upregulated PKMYT1 expression by interacting with the DNA binding region of PKMYT1 in GBC cells, thereby promoting the phosphorylation of CDK1 and inducing Gem-R. Treatment with a combination of the PKMYT1 shRNA and gemcitabine significantly reduced the growth of GBC cells induced by overexpression of ELF3 in vitro and in vivo.
Conclusions:
ELF3/PKMYT1/CDK1 axis significantly regulates Gem-R to GBC cells and may represent a promising drug target for treating GBC patients.
Insights
ELF3 drives gemcitabine resistance in gallbladder cancer (GBC) by upregulating PKMYT1 and CDK1. Targeting the ELF3/PKMYT1/CDK1 axis offers a promising strategy to overcome gemcitabine resistance in GBC patients.
Area of Science:
- Molecular Oncology
- Cancer Therapeutics
- Genomics and Proteomics
Background:
- Gemcitabine is the standard chemotherapy for gallbladder cancer (GBC).
- Development of gemcitabine resistance (Gem-R) limits treatment efficacy in GBC.
- Molecular mechanisms underlying Gem-R in GBC remain largely unknown.
Purpose of the Study:
- To investigate the role of ELF3 in gemcitabine resistance in GBC.
- To elucidate the underlying molecular mechanisms of ELF3-mediated Gem-R.
Main Methods:
- RNA sequencing to identify genes associated with Gem-R.
- Analysis of ELF3 expression in GBC datasets and cell lines.
- In vitro and in vivo assays (e.g., CCK-8, xenograft models) to assess gemcitabine sensitivity and ELF3 function.
- Luciferase reporter assays to determine ELF3's interaction with the PKMYT1 promoter.
Main Results:
- Elevated ELF3 expression positively correlates with Gem-R in GBC.
- ELF3 overexpression promotes Gem-R, while ELF3 knockdown enhances gemcitabine sensitivity.
- ELF3 upregulates PKMYT1, leading to CDK1 phosphorylation and Gem-R; combination therapy with PKMYT1 shRNA and gemcitabine inhibits GBC growth.
Conclusions:
- The ELF3/PKMYT1/CDK1 axis is a key regulator of gemcitabine resistance in GBC.
- This axis represents a potential therapeutic target for overcoming Gem-R in GBC patients.
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