Effect of miR-218-2-3P on proliferation and apoptosis of NK/T-cell lymphoma by targeting SIN3A / 上海交通大学学报（医学版）

ABSTRACT

Objective • To investigate the expression of miR-218-2-3P in NK/T-cell lymphoma, and the effect of miR-218-2-3P on the proliferation, apoptosis and cycle of NK/T-cell lymphoma by targeting SIN3A. Methods • Quantitative real-time PCR (qPCR) was used to detect the expressions of miR-218-2-3P in normal NK cells and NK/T-cell lymphoma cells NK92MI and NKYS. Lipofectamine 3000 was used to transfect the inhibitor containing nonsense sequences (inhibitors NC), miR-218-2-3P inhibitor and the same dose of transfection reagent without any fragment into NK92MI cells, which were divided into three groups. qPCR and Western blotting were used to detect the expression levels of miR-218-2-3P and SIN3A protein in the inhibitor NC group, the miR-218-2-3P inhibitor group and the blank control group, respectively. The cell proliferation activities of the three group were measured by CCK8 method. The apoptosis rates and cell cycles of the three group were determined by flow cytometry. The double luciferase reporter gene assay was performed to detect whether SIN3A was a target gene of miR-218-2-3P. NK92MI cells were transfected with miR-218-2-3P inhibitor+SIN3A small interfering RNA (si-SIN3A) and miR-218-2-3P inhibitor+nonsense sequences small interfering RNA (si-NC), respectively, which were divided into two groups. The cell proliferation activities of the two groups were detected by CCK8 method. Results • Compared with the normal NK cells, the expressions of miR-218-2-3P in NK92MI and NKYS cells significantly increased (both P<0.05). Compared with the blank control group and the inhibitor NC group, the proliferation activity of NK92MI cells in the miR-218-2-3P inhibitor group decreased (both P<0.05), the apoptosis rate increased (both P<0.05) and cell cycle arrest occurred in G0/G1 phase (both P<0.05). The double luciferase reporter gene assay showed that SIN3A was the target gene of miR-218-2- 3P. Compared with the blank control group and the inhibitor NC group, the SIN3A protein expression of the miR-218-2-3P inhibitor group was increased (both P<0.05). The down-regulation of SIN3A expression could restore the proliferation activity of cells weakened by miR-218-2-3P inhibitor (both P<0.05). Conclusion • miR-218-2-3P is highly expressed in the NK92MI and NKYS cell lines. miR-218-2-3P may affect the proliferation, apoptosis and cycle of NK/T-cell lymphoma through targeted regulation of SIN3A.