Analgesic effects of miR-362 on bone cancer pain process via targeting HDAC6 in rats / 西安交通大学学报(医学版)

ABSTRACT

【Objective】 To detect the expressions of microRNA（miR）-362 and histone deacetylase 6 （HDAC6） in bone cancer pain （BCP） rats and investigate the analgesic effect of miR-362 and its potential analgesic mechanism. 【Methods】 The BCP model was developed by injecting Walker 256 mammary gland carcinoma cells into bone marrow cavity. Plasmid transfection was used to regulate the expressions of miR-362 and HDAC6. The Van Frey filaments and radiant heat instrument were used to detect the paw withdrawal threshold （PWT） and paw withdrawal latency （PWL）. qRT-PCR was used to detect mRNA expression levels of miR-362 and HDAC6， and Western blotting was used to detect protein expression of HDAC6 and nuclear factor kappa-B p65 （NF-κB p65）. ELISA assay was used to detect the protein levels of interleukin （IL）-1β， IL-6 and tumor necrosis factor α （TNF-α）. Luciferase activity assay was used to determine the relationship between miR-362 and HDAC6. 【Results】 Compared to sham group， the significant decrease of PWT and PWL， decrease of miR-362 and the increase of HDAC6 mRNA and protein in the spinal were detected in BCP group （P<0.05）. Compared to BCP group， the significantly increase of PWT and PWL and decrease of HDAC6 mRNA and protein in the spinal were detected in BCP+LV-miR-362 group （P<0.05）. Compared to BCP+LV-miR-362 group， PWT and PWL significantly decreased in BCP+LV-miR-362+LV-HDAC6 group （P<0.05）. In addition， compared to BCP group， significant decreases of NF-κB p65， IL-1β， IL-6 and TNF-α in spinal were detected in BCP+LV-HDAC6 siRNA group （P<0.05）. Moreover， compared to mimic miR-362+HDAC6 3’UTRMUT group， the luciferase activity significantly decreased in mimic miR-362+HDAC6 3’UTRWT group （P<0.05）. 【Conclusion】 As a key factor regulating the mechanism of BCP through “HDAC6-NF-κB p65” signal pathway in rats， targeting miR-362 may be a novel therapeutic method for BCP.