RESUMEN
Objective:To investigate the mechanism underlying bone metastasis pain in lung cancer mediated by Gm31083/miR-450b-5p/EphB1 pathway.Methods:Between January 2020 and December 2021, a total of 20 healthy specific pathogen-free (SPF) grade Sprague-Dawley mice were selected and randomly divided into a sham operation group and a model group using a random number table method, with 10 mice in each group. A lung cancer bone metastasis pain model was established by injecting human lung adenocarcinoma A549 cells into the femur of each mouse in the model group. The middle part of the left plantar area of the mouse was stimulated with 2 g Von Frey fine fibers, and the mouse's foot retraction response was observed for 5-6 seconds. The pain threshold of each mouse was measured using a hot plate instrument. The expression of miR-450b-5p was determined by real-time fluorescence quantitative polymerase chain reaction. The expression of Gm31083 and EphB1 proteins was determined by western blot assay.Results:The foot contraction response rate in the model group was (31.98 ± 6.36)%, which was significantly higher than (22.78 ± 4.54)% in the sham operation group ( t = -3.72, P < 0.05). The heat pain threshold in the model group was (8.18 ± 2.53) seconds, which was significantly lower than (15.42 ± 3.97) seconds in the sham operation group ( t = 4.86, P < 0.001). The relative expression level of miR-450b-5p in the model group was (1.62 ± 0.29), which was significantly higher than (1.00 ± 0.04) in the sham operation group ( t = -6.70, P < 0.001). The grayscale values of Gm31083 and EphB1 proteins in the model group mice were (1.23 ± 0.21) and (2.73 ± 0.28), respectively, which were significantly higher than (0.67 ± 0.18) and (1.25 ± 0.24) in the sham operation group ( t = -6.40, -12.69, both P < 0.001). Conclusion:Gm31083/miR-450b-5p/EphB1 is highly expressed in mice suffering from bone metastasis pain after developing lung cancer, and may become a new biomarker for evaluating bone metastasis pain in lung cancer.
RESUMEN
Objective: To investigate the expression, clinical significance and biological function of miR-450b-5p in hepatocellular carcinoma (HCC) so as to make a preliminary exploration of the mechanism in HCC. Methods: Real-time PCR was used to detect miR-450b-5p expression in HCC tissues and cells. Statistical analysis was employed to explore the correlations of miR-450b-5p with clinicopathologic features and prognosis. And Cox regression analysis was applied to analyze the relationship between clinicopathologic characteristics and prognosis. Transwell assay and MTT assay were used to study the effects of miR-450b-5p on migration, invasion and proliferation of HCC cells. Bioinformatics tools were applied to predict the potential target gene of miR-450b-5p. Luciferase reporter assay and Western blot were employed to determine the correlation between miR-450b-5p and its target gene. Results: miR-450b-5p was significantly down-regulated in HCC tissues and cell lines (P<0.001). miR-450b-5p expression was significantly correlated with tumor size (P=0.026), portal vein infiltration (P=0.013), TNM stage (P=0.020), and the prognosis of HCC patients. Cox regression analysis showed that miR-513a-5p expression was an independent prognostic factor for the HCC patients' survival. Overexpressed miR-450b-5p notably inhibited migration, invasion and proliferation of MHCC-97H cells, while miR-450b-5p inhibitors had the opposite effects on Hep3B cells. Bioinformatics analysis suggested that Sex Determining Region Y Box 2 (SOX2) might be the target of miR-450b-5p. Furthermore, luciferase reporter assay indicated that the luciferase activity of SOX2-3'-UTR was negatively regulated by miR-450b-5p. Consistently, Western blot showed that miR-450b-5p negatively regulated the expression of SOX2 in HCC cells. Conclusion: miR-450b-5p suppresses migration, invasion and proliferation of HCC cells by targeting SOX2.