Comparison of expression levels of hsa-miR-155, INPP5D and IFN-γ level in NK cells isolated from refractory/relapsed or responsive patients with large diffuse B-cell lymphoma (DLBCL) and healthy donors

Introduction and Objectives: The most common high-grade form of non-hodgkin lymphoma (NHL) is diffuse large B-cell lymphoma (DLBCL). NK cells stimulated by CD16 or IL-12 and IL-18 induces the miR-155 overexpression. Has-miR-155 overexpression targets the Src Homology 2 Domain-containing Inositol 5′-Phosphatase (SHIP-1) as a negative regulator, consequently enhances IFN-γ production. IFN-γ is a critical cytokine for the tumor surveillance, therefore understanding the molecular pathways of IFN-γ expression lead to identifying the potential therapeutic targets for cancer. Therefore, we evaluated whether the expression levels of hsa-miR-155-5p, INPP5D and IFN-γ level are different in the NK cells from patients with DLBCL in comparison with healthy donors.Materials and Methods: Patients were consecutively selected from Seyed-al-Shohada and Alzahra Hospital in Isfahan, Iran. Peripheral blood was obtained from healthy people (n=12) and responsive patients with DLBCL (n=10) or refractory/relapsed patients with DLBCL (n=12). Peripheral blood mononuclear cells (PBMCs) (Ficoll-Hypaque gradient) and then NK cells (NK cell isolation kit, Miltenyi Biotec, Bergisch Gladbach, Germany) were isolated. Levels of hsa-miR-155-5p and INPP5D were analyzed by qRT-PCR. The concentrations of IFN-γ in the culture supernatant were measured with the Human IFN-γ ELISA MAX™ Deluxe (Bio-Legend, San Diego, CA) after 72 hours.Results: We observed the increased expression level of hsa-miR-155-5p (P=0.01 LSD Post-Hoc) and decreased expression level of INPP5D (P=0.017 and P=0.001 LSD Post-Hoc) in healthy donors relative to responsive patients or refractory/relapsed patients with DLBCL. The IFN-γ concentration in the culture supernatants of NK cells from healthy donors was more than responsive patients and refractory/relapsed patients (P=0.05 and P=0.001, LSD Post-Hoc).Conclusion: Decreased expression of hsa-miR-155-5p and IFN-γ might be associated with low NK cell function and impaired response to treatment in patients with DLBCL. Therefore, it is necessary to further evaluate the miR-155-5p- INPP5D- IFN-γ pathway.

کلیدواژه ها:

DLBCL ، hsa-miR-155-5p ، INPP5D ، IFN-γ ، NK cell