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Updated: Jan 15, 2026
High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
Published on: August 3, 2011
Utility of peritoneal fluid for multiplexed mRNA profiling of peritoneal metastasis from pancreatic cancer
Sharuya Kulatheivam1, Lea Lecanda Mariager Jakobsen1, Mark Burton2
1Department of Pathology, Odense University Hospital, Odense, Denmark; Odense Pancreas Center (OPAC) and Odense PIPAC Center (OPC), Odense University Hospital, Odense, Denmark.
Background:
Patients with peritoneal metastasis (PM) have poor survival rates, and few methods for prognostic stratification of patients with proven PM exist. Peritoneal fluid (PF) provides a direct reflection of the microenvironment of the peritoneal cavity, and transcriptomic profiling of PF represents a novel and potentially clinically valuable approach that warrants investigation. We aimed to evaluate the utility of formalin-fixed paraffin embedded (FFPE) PF sediments left over after conventional peritoneal cytology diagnosis for multiplexed mRNA expression profiling and to identify dysregulated mRNAs in PM vs. controls. Furthermore, we explored whether certain mRNAs upregulated in PM hold prognostic value.
Methods:
Samples included were FFPE sediments of malignant PFs from cases of PM from pancreatic cancer (PM-PC, n = 19) scheduled for pressurized intraperitoneal aerosol chemotherapy (PIPAC) and benign ascites (controls, n = 16). RNA was extracted from the FFPE sediments. Expression profiling of 760 cancer-related mRNAs was performed, followed by unsupervised clustering, differential gene expression analysis, ROC/AUC analysis, generation of KM plots, and Cox proportional hazards regression. Cellular expression of proteins encoded by selected dysregulated mRNAs was analyzed using immunocytochemistry.
Results:
In PF sediments with malignant cells, 56 upregulated and 130 downregulated mRNAs were identified. Selected up- and downregulated mRNAs were similarly dysregulated at the protein level. Upregulation of six mRNAs (ESRP2, EPCAM, SFN, ITGB6, ESRP1, and GRHL2) was associated with short-term survival (P < 0.0025).
Conclusion:
Our data indicate that PF from PM-PC patients is suitable for transcriptomic analysis. Numerous upregulated mRNAs known to play a role in tumour progression were identified. Our data indicate that mRNA profiling may be a clinically useful tool for further prognostic stratification of patients with proven PM, but larger studies are needed to validate our findings.
Insights
Peritoneal fluid mRNA profiling can identify cancer markers and aid in prognostic stratification for patients with peritoneal metastasis (PM). This novel approach uses leftover cytology samples for transcriptomic analysis, offering new insights into cancer progression.
Area of Science:
- Oncology
- Molecular Biology
- Genomics
Background:
- Peritoneal metastasis (PM) is associated with poor patient survival.
- Limited prognostic stratification methods exist for PM patients.
- Peritoneal fluid (PF) offers a direct view of the peritoneal cavity's microenvironment.
Purpose of the Study:
- To assess the utility of formalin-fixed paraffin-embedded (FFPE) PF sediments for mRNA expression profiling.
- To identify messenger RNAs (mRNAs) dysregulated in PM compared to controls.
- To explore the prognostic value of upregulated mRNAs in PM.
Main Methods:
- RNA extraction from FFPE PF sediments of pancreatic cancer PM (n=19) and benign ascites (n=16).
- Multiplexed profiling of 760 cancer-related mRNAs.
- Bioinformatic analyses including differential gene expression, survival analysis (Kaplan-Meier, Cox regression), and immunocytochemistry.
Main Results:
- 56 upregulated and 130 downregulated mRNAs identified in malignant PF sediments.
- Dysregulation of selected mRNAs was confirmed at the protein level.
- Upregulation of six specific mRNAs (ESRP2, EPCAM, SFN, ITGB6, ESRP1, GRHL2) correlated with shorter survival.
Conclusions:
- FFPE PF sediments are suitable for transcriptomic analysis.
- Numerous tumor-progression-associated mRNAs are upregulated in PM.
- mRNA profiling shows potential as a clinical tool for prognostic stratification in PM patients, pending larger validation studies.
