Diospyros Peregrina Fruit Preparation Mediated Immunomodulation of Lymphocytes Isolated From The Blood of Breast Cancer Patients.

BACKGROUND

Breast cancer is an uncontrolled growth of epithelial cells. The loss of Breast Cancer gene1 (BRCA1) activity due to mutation or down-regulation of gene expression promotes tumorigenesis and increases the risk of breast cancer.

OBJECTIVES

We aimed to pulsate lymphocytes of breast cancer patients and normal individuals, using Diospyros peregrina fruit preparation (DFP) to study the cancer-protective immunity, and the signal transduction processes involved with it. We also investigated the role of DFP in the release of lymphocytic nitric oxide (NO), which is a key tumoricidal agent, known to regulate T-cell proliferation, cytokine production, cell signaling, and apoptosis.

METHODS

Using Ficoll-Hypaque gradient centrifugation, lymphocytes were isolated from the blood of 12 patients and 12 normal individuals. Cells were treated with or without DFP (2.5 µg/ml) for 48 hours. Both non-stimulated and stimulated cells were then subjected to MTT and NO release assay; following which qPCR was performed to estimate mRNA levels and percentage enrichment of certain genes.

RESULTS

DFP stimulates lymphocytic proliferation (p=0.0118) and release of NO (p=0.01) significantly. DFP also noticeably enhances the expression of T helper (TH) cell 1 specific interferon-gamma (IFNG), interleukin 12 (IL12), T-Box Transcription Factor 21 (TBX21) and signal transducer and activator of transcription 1 (STAT1) genes. DFP treatment significantly increases tumor protective immunity by decreasing the expression levels of TH2 network-specific GATA3 and interleukin 4 (IL4) genes but increasing the expression levels of TH1 network-specific IFNG, IL12, TBX21, and STAT1 genes.

CONCLUSION

DFP increases the expression levels of TH1 specific network genes which in turn help in evoking tumor protective immunity.