ABSTRACT
To investigate the effect of miR-30a/HMGA2-mediated autophagy in osteosarcoma cells on apoptosis induced by chemotherapeutics. Methods: A total of 30 osteosarcoma tissues of sensitive and resistant to chemotherapeutics were divided into a chemotherapy-sensitive group and a chemotherapy-resistant group. The mRNA expression levels of miR-30a and high mobility group protein A2 (HMGA2) in the chemotherapy-sensitive group and the chemotherapy-resistant group, and the mRNA expression levels of miR-30a in osteosarcoma U2-OS cells treated by cisplatin, doxorubicin and methotrexate at different concentrations were detected by real-time PCR. The expression levels of autophagy related protein Beclin 1, microtubule associated protein 1 light chain 3B (LC3B) and autophagy factor P62 were detected by Western blotting. The osteosarcoma U2-OS cells were transfected with miR-30a mimics and miR-30a inhibitors to construct a miR-30a high expression group, a miR-30a low expression group and a control group. The expression levels of Beclin 1, LC3B and P62 in osteosarcoma U2-OS cells after treatment of cisplatin and doxorubicin in these 3 groups were detected by Western blotting; the level of autophagy was detected by monodansylcada (MDC) staining; the level of ROS was detected by dihydroethidium (DHE); the level of cell surviving rate was detected by cell counting kit-8 (CCK-8); the level of apoptosis was detected by annexin APC/PI double staining; the level of mitochondria oxidative damage was detected by mitochondrial membrane potential assay kit with JC-1 (JC-1 method). The interaction between miR-30a and HMGA2 was detected by dual luciferase reporter assay. The osteosarcoma U2-OS cells were transfected with HMGA2 mimics and HMGA2-shRNA to construct a high HMGA2 group, a low HMGA2 group, and a control group. The expression levels of Beclin 1, LC3B and P62 in osteosarcoma U2-OS cells after the treatment of cisplatin were detected by Western blotting. Results: The level of miR-30a in the chemotherapy-resistant tissues was significantly lower than that in the chemotherapy-sensitive tissues (P<0.05), and the expression of HMGA2 was opposite comparing to that of miR-30a (P<0.05). After the treatment by low concentration (5 μmol/L) of chemotherapeutics, the level of miR-30a was down-regulated in osteosarcoma U2-OS cells, accompanied with up-regulation of Beclin 1 and LC3B (P<0.01) and down-regulation of P62 (P<0.01). Compared with the control group, the expression levels of Beclin 1 and LC3B were significantly decreased (P<0.05), and the expression level of P62 was significantly increased (P<0.05) in the miR-30a high expression group, which was opposite in the miR-30a low expression group. In the miR-30a high expression group treated by chemotherapeutics, the level of autophagy and the cell survival rate were lower than those in group with low expression of miR-30a, while the levels of ROS, the mitochondrial oxidative damage and the apoptosis were higher than those in group with low expression of miR-30a (all P<0.05). The targeting interaction between HMGA2 and miR-30a were verified by dual luciferase reporter assay. Compared with the control group, the expression levels of Beclin 1 and LC3B were significantly increased (P<0.05), and the expression level of P62 was significantly decreased (P<0.05) in the HMGA2 high expression group, which was opposite in the HMGA2 low expression group. Conclusion: Suppression of miR-30a/HMGA2-mediated autophagy in osteosarcoma cells is likely to enhance the therapeutic effect of chemotherapeutics.
Subject(s)
Humans , Apoptosis , Apoptosis Regulatory Proteins , Autophagy , Beclin-1 , Bone Neoplasms , Cell Line, Tumor , HMGA2 Protein , Metabolism , MicroRNAs , Genetics , OsteosarcomaABSTRACT
Objective To study the biomechanical properties of porous titanium cages used for different lumbar interbody fusion surgeries. Methods The three-dimensional (3D) finite element model of the lumbar spine was constructed, and mechanical parameters of porous materials were obtained by mechanical test. The biomechanical properties of porous titanium cages in anterior lumbar interbody fusion (ALIF), posterior lumbar interbody fusion (PLIF), transforaminal lumbar interbody fusion (TLIF), direct lateral interbody fusion (DLIF) were compared. Results After lumbar interbody surgery, the predicted range of motion (ROM) and the maximum stress in cage of DLIF model and ALIF model were substantially lower than those of PLIF model and TLIF model. The maximum stress in endplate of DLIF model, ALIF model and TLIF model were obviously lower than that of PLIF model. Conclusions DLIF with the porous cage showed advantages in biomechanical properties, which was simple to operate and suitable for minimally invasive surgery in clinical practice. DLIF performed the superior comprehensive properties.
ABSTRACT
Objective To investigate the clinical efficacy and safety of testosterone supplement and Mreceptor blockers in the treatment of patients with lower urinary tract symptoms(LUTS)in late -onset hypogonadism(LOH). Methods 28 cases diagnosed as LOH with mild to moderate LUTS were collected.They were given testosterone supplementation (oral testosterone undecanoate capsules,80mg,2 times/d)and M receptor blocker (oral succinate solifenacin tablet,5mg,1 time /d)treatment for 1 -3 months.After treatment for 1 month and 3 months respectively, reviewd PSA,serum total testosterone (TT),international prostate of urinary storage symptoms score (urinary storage IPSS),quality of life (QOL)score,international index of erectile function (IIEF -5)score,maximum urinary flow rate (Qmax),residual urine (Ru)and rectal examination (DRE).The improvement of LUTS before and after treatment was evaluated.Results 1 month after treatment,1 patient was difficult to tolerate the solifenacin side effects and took testosterone supplementation alone.The rest 27 patients were able to take medicine for 3 months.After 1 month and 3 months treatment,the IPSS score had significant differences compared with before treatment[(10.3 ±2.1)points vs (14.2 ±3.3)points and (9.42 ±1.8)points vs (14.2 ±3.3)points,t =13.67,14.72,all P <0.05 ].After 3 months treatment,the QOL and IIEF -5 scores were (2.1 ±0.7)points vs (4.3 ±0.6)points and (16.8 ± 3.6)points vs (11.9 ±2.5)points,Qmax was (12.5 ±5.6)mL/s vs (9.8 ±4.8)mL/s(t =6.42,5.64,14.92,all P <0.05 ),the difference was statistically significant compared with before treatment.There were no significant changes in PSA and RU before and after treatment.All patients had no severe complications such as acute urinary retention.Conclusion Combination of testosterone and testosterone in the treatment of LUTS patients in LOH is safe and effective,and can significantly improve the LUTS and quality of life.