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.
ABSTRACT
Objective To explore miR-362-3 p expression in laryngeal cancer tissues and its effect on migration of Hep2 cells. Methods miR-362-3 p expression in 50 pairs of tumor and adjacent normal tissues was detected by real-time PCR after sample collection. The relationships between miR-362-3 p expression and clinical pathological characteristics in patients with laryngeal cancer were analyzed.mi R-362-3 p mimic, inhibitor, and control microRNA were transfected into Hep-2 cells. Transfection efficiency was determined by real-time PCR. Wound-healing and transwell assays were used to evaluate Hep-2 migration. Results miR-362-3 p expression was significantly higher in cancer tissues than in adjacent normal tissues (P ＜ 0.05). miR-362-3 p expression was statistically significantly related to node metastasis and clinical stage. Transfection with miR-362-3 p mimic and inhibitor significantly increased and decreased Hep-2 cell migration, respectively (both P ＜ 0.05). Conclusion miR-362-3 p is up-regulated in laryngeal cancer tissues and promotes laryngeal cancer cell migration, suggesting that it acts as a potential oncogene in laryngeal cancer.
ABSTRACT
PURPOSE: Colorectal cancer (CRC) is the third most common cancer in China and poses high morbidity and mortality. In recent years, increasing evidence has indicated that microRNAs played important functions in the occurrence and development of tumors. The purpose of this study was to identify the biological mechanisms of miR-362 in CRC. MATERIALS AND METHODS: Quantitative real-time PCR was carried out to assess the expression of miR-362 and SIX1. The Kaplan-Meier method was employed to evaluate the 5-year overall survival of CRC patients. The proliferative and invasive abilities of CRC cells were assessed by MTT and transwell assays. RESULTS: miR-362 was significantly decreased in CRC tissues and cell lines, compared to the normal tissues and normal cells. A significant connection was confirmed between the overall survival of 53 CRC patients and low expression of miR-362. Downregulation of miR-362 inhibited the proliferation and invasion through binding to the 3′-UTR of SIX1 mRNA in CRC. Additionally, we discovered that SIX1 was a direct target gene of miR-362 and that the expression of miR-362 had a negative connection with SIX1 expression in CRC. SIX1 could reverse partial functions in the proliferation and invasion in CRC cells. CONCLUSION: miR-362 may be a prognostic marker in CRC and suppress CRC cell proliferation and invasion in part through targeting the 3′-UTR of SIX1 mRNA. The newly identified miR-362/SIX1 axis provides insight into the progression of CRC.