# Specific Marker Gene Analysis for Primary Central Nervous System Lymphoma Based on Methylation Difference and Development of Detection Primers

## TL;DR

This study identifies RHEB gene methylation as a potential biomarker for PCNSL and develops primers to detect it in blood samples.

## Contribution

Development of methylation-specific primers targeting RHEB for PCNSL detection in plasma.

## Key findings

- Promoter hypermethylation of RHEB gene is a specific biomarker for PCNSL.
- Methylation primers targeting RHEB can distinguish PCNSL from other CNS diseases.
- Primer sensitivity was validated through in vitro methylation and demethylation experiments.

## Abstract

Primary central nervous system lymphoma (PCNSL) is a central nervous system disease with high mortality, wide variation in symptoms, and difficult diagnosis. The development of molecular markers for PCNSL is still in its initial stages.

The databases were analyzed by 450 K and RRBS, and the differential gene structure was displayed through Venn diagram, heatmap, and enrichment. ROC and survival curve analyses were performed on the marker genes. The specificity of PCNSL in blood samples of patients was verified by qPCR, and the sensitivity of the detection primers was verified by in vitro methylation and demethylation drug treatment cell experiments.

Twenty‐six sites were identified using 450 K microarray analysis. The RRBS analysis team found 14,867 sites. The methylation site located in the promoter region of REHB was verified, and the results showed that the primers for this site were able to distinguish patients with PCNSL and were sensitive to the detection of methylation levels.

A pair of methylation primers targeting the RHEB promoter region were obtained, which demonstrated the potential to distinguish patients with PCNSL from those with other CNS diseases. These findings should be considered preliminary and serve as proof‐of‐concept for further validation in larger cohorts.

Promoter hypermethylation of the RHEB gene is identified as a specific biomarker for Primary Central Nervous System Lymphoma (PCNSL). This study develops methylation‐specific primers, demonstrating their power to distinguish PCNSL from other CNS diseases via a novel plasma‐based detection method.