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Ribonucleotide reductase M2 subunit silencing suppresses tumorigenesis in pancreatic cancer via inactivation of
Jinlan Shan1, Zhen Wang2, Qiuping Mo3
1Department of Surgery, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China; Department of Cancer Institute, Zhejiang University, Hangzhou, Zhejiang, China.
Background/Objectives:
Ribonucleotide Reductase M2 subunit (RRM2) is elevated in pancreatic cancer and involved in DNA synthesis and cell proliferation. But its specific mechanism including genetic differences and upstream regulatory pathways remains unclear.
Methods:
We analyzed RRM2 expression of 178 pancreatic cancer patients in Gene Expression Profiling Interactive Analysis (GEPIA) database. Besides, more pancreatic cancer specimens were collected and detected RRM2 expression by immunohistochemistry. RRM2 knockdown by shRNA was applied for functional and mechanism analysis in vitro. Xenograft tumor growth was significantly slower by RRM2 silencing in vivo.
Results:
It showed that high RRM2 expression had a poorer overall survival and disease free survival. RRM2 expression was higher in tumor grade 2 and 3 than grade 1. Immunohistochemistry data validated that high RRM2 expression predicted worse survival. RRM2 knockdown significantly reduced cell proliferation, inhibited colony formation and suppressed cell cycle progress. Further mechanism assay showed silencing RRM2 lead to inactivation of PI3K/AKT/mTOR pathway and inhibition of mutant p53, which induce S phase arrest and/or apoptosis. In panc-1 cells, S-phase arrest mediated by mutant p53 inhibition, p21 increase and cell cycle related proteins change. While in miapaca-2 cells, induction of apoptosis and S-phase arrest mediated by CDK1 played a coordinated role.
Conclusion:
Taken together, high RRM2 expression was associated with worse prognosis. Importantly, RRM2 knockdown deactivated PI3K/AKT/mTOR pathway, resulting in cell cycle arrest and/or apoptosis. This study shed light on the molecular mechanism of RRM2 in pancreatic tumor progression and is expected to provide a new theoretical basis for pancreatic cancer treatment.
Insights
High Ribonucleotide Reductase M2 subunit (RRM2) expression in pancreatic cancer correlates with poor prognosis. Silencing RRM2 inhibits tumor growth by deactivating the PI3K/AKT/mTOR pathway, leading to cell cycle arrest and apoptosis.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Ribonucleotide Reductase M2 subunit (RRM2) is implicated in pancreatic cancer progression.
- The precise molecular mechanisms and regulatory pathways of RRM2 in pancreatic cancer remain largely undefined.
Purpose of the Study:
- To investigate the prognostic value of RRM2 in pancreatic cancer.
- To elucidate the underlying molecular mechanisms of RRM2 in pancreatic tumor growth and progression.
Main Methods:
- Analysis of RRM2 expression in 178 pancreatic cancer patients using the GEPIA database.
- Immunohistochemical validation of RRM2 expression in pancreatic cancer specimens.
- In vitro RRM2 knockdown using shRNA to assess functional impact and mechanistic pathways.
- In vivo xenograft studies to evaluate the effect of RRM2 silencing on tumor growth.
Main Results:
- Elevated RRM2 expression was significantly associated with poorer overall and disease-free survival, and higher tumor grade.
- RRM2 knockdown markedly reduced pancreatic cancer cell proliferation, colony formation, and induced cell cycle arrest.
- Silencing RRM2 led to the inactivation of the PI3K/AKT/mTOR pathway and inhibition of mutant p53, triggering S-phase arrest and/or apoptosis.
Conclusions:
- High RRM2 expression serves as a biomarker for poor prognosis in pancreatic cancer.
- Targeting RRM2 by knockdown deactivates the PI3K/AKT/mTOR pathway, inducing cell cycle arrest and apoptosis, offering a potential therapeutic strategy for pancreatic cancer.
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