MiRNA-138-5p: A strong tumor suppressor targeting PD-L-1 inhibits proliferation and motility of breast cancer cells and induces apoptosis.

Abstract

MicroRNAs are important regulators in multiple cellular processes and are closely related to a variety of cancers including breast cancer (BC). Immunotherapy using different methods such as modulating immune check points has been known as an advanced and successful procedure in cancer treatment. Here we investigated the effects of miRNA-138-5p restoring on Programmed Death Ligand 1 (PD-L-1) expression, BC biological behaviors and T-cell exhaustion. Breast cancer specimens and cell lines were provided and qRT-PCR and western blotting were used to measure the expression of miRNA-138-5p, PD-L-1 and other underlying genes. MTT and colony formation assays and scratch test were employed to specify proliferation, cloning and migration in miRNA-138-5p-transfected MDA-MB-231\xa0cells respectively. DAPI staining assay and flow-cytometry were used to investigate apoptosis rate and cell cycle development. Finally, isolated T-cells were co-cultured with transfected BC cells to explore the effect of miRNA-138-5p on T-cell exhaustion. qRT-PCR revealed down-regulation ofmiRNA-138-5p conversely, up-regulation of PD-L-1 in BC tissues and cell lines. Transfection of miRNA-138-5p into MDA-MB-231\xa0cells inhibited PD-L-1 expression. Western blotting, MTT and colony formation assays affirmed the anti-proliferative effect ofmiRNA-138-5p through down-regulating PI3K/AKT pathway. Also, miRNA-138-5p induced apoptosis in BC cells via up-regulating Caspase-9 and Caspase-3 and arresting cell cycle at sub-G1 phase. Moreover, scratch test and western blotting indicated that miRNA-138-5p inhibits cell motility via targeting MMP2, MMP9 and vimentin but up-regulating E-cadherin. Finally, miRNA-138-5p restrains T-cell exhaustion via suppressing PD-L-1 expression in BC cells leading to disrupt PD-L-1/PD-1 interaction and modulate effector cytokines in T-cells.