ABSTRACT
Post-translational modification of proteins is involved in the occurrence of endometriosis (EM); however, the role of ubiquitination modification in EM remains unclear. Integrin ß3 (ITGB3) is one of the ß-subunits of integrins, which plays a key role in tumor progression. In this study, we investigated the roles of ITGB3 and ITCH, one of the ubiquitin E3 ligases, in ectopic endometrial stromal cells (ESCs) and EM. Primary ectopic ESCs and normal ESCs were isolated and purified. Western blot was used to detect the expression of ITGB3 and ITCH in ESCs. The interaction between ITGB3 and ITCH in ESCs was investigated by the co-immunoprecipitation and ubiquitylation analysis. With or without the overexpression of ITCH and/or ITGB3, the proliferation and invasion of ectopic ESCs were detected by the CCK8 assay and transwell migration assay, respectively. We found that ITGB3 is upregulated in ectopic ESCs from patients with EM. ITCH interacts with ITGB3 by co-immunoprecipitation, and ITCH-overexpressing significantly increased the ubiquitination of ITGB3. The data of the CCK8 assays showed that ITGB3 overexpression significantly promoted cell proliferation of ectopic ESCs at 12, 24, 48, and 72 h. The transwell migration assays showed that ITGB3 overexpression significantly enhanced the invasive ability. However, ITCH had the opposite effects in both assays. Our findings indicate that ITCH-mediated ubiquitylation of ITGB3 regulates the proliferation and invasion ability of ectopic ESCs in EM.
ABSTRACT
PURPOSE: The purpose of this study was to observe the effects of micro ribonucleic acid (miR)-505-5p on the proliferation and apoptosis of osteosarcoma cells, and to further investigate its potential mechanism. METHODS: Human osteosarcoma U2-OS cell lines were divided into Control group, miR-505-5p nonsense sequence (NS) group and miR-505-5p inhibitor group. Subsequently, cell proliferation and apoptosis in each group were observed. Finally, the effect of miR-505-5p on the in vivo growth of osteosarcoma was explored by means of subcutaneous tumor formation assay. RESULTS: The expression of miR-505-5p in the cancer tissues was remarkably higher than in normal paracancer tissues of osteosarcoma patients. U2-OS cell lines cultured in vitro in miR-505-5p inhibitor group manifested notably weakened proliferative ability after transfection with miR-505-5p inhibitor. Colony formation assay showed that the number of colonies formed in miR-505-5p inhibitor group was obviously smaller than that in Control group. The results of Western blotting assay indicated that the inhibition of miR-505-5p markedly increased the expression of Bax and decreased Bcl-2 in cancer cells (p<0.05). Furthermore, it was revealed that the inhibited miR-505-5p could distinctly up-regulate the protein expression level of RASSF8 in cancer cells. Furthermore, the miR-505-5p inhibition was able to prominently repress the subcutaneous tumor formation ability of osteosarcoma cells. CONCLUSIONS: The expression level of miR-505-5p is raised significantly in the cancer tissues of osteosarcoma patients, and the inhibition of miR-505-5p can up-regulate RASSF8 to suppress the proliferation and promote the apoptosis of osteosarcoma cells.
Subject(s)
Biomarkers, Tumor/metabolism , MicroRNAs/metabolism , Osteosarcoma/genetics , Tumor Suppressor Proteins/metabolism , Apoptosis , Cell Line, Tumor , Cell Proliferation , Female , Humans , Male , Osteosarcoma/pathologyABSTRACT
OBJECTIVE: To evaluate the clinical effect of the minimally invasive transforaminal lumbar interbody fusion combined with posterolateral fusion and unilateral fixation using a tubular retractor in the management of degenerative lumbar disease. METHODS: A retrospective analysis was conducted to analyze the clinical outcome of 58 degenerative lumbar disease patients who were treated with minimally invasive transforaminal lumbar interbody fusion combined with posterolateral fusion and unilateral fixation during December 2012 to January 2015. The spine was unilaterally approached through a 3.0-cm skin incision centered on the disc space, located 2.5 cm lateral to the midline, and the multifidus muscles and longissimus dorsi were stripped off. After transforaminal lumbar interbody fusion and posterolateral fusion the unilateral pedicle screw fixation was performed. The visual analogue scale (VAS) for back and leg pain, the Oswestry disability index (ODI), and the MacNab score were applied to evaluate clinical effects. The operation time, peri-operative bleeding, postoperative time in bed, hospitalization costs, and the change in the intervertebral height were analyzed. Radiological fusion based on the Bridwell grading system was also assessed at the last follow-up. The quality of life of the patients before and after the operation was assessed using the short form-36 scale (SF-36). RESULTS: Fifty-eight operations were successfully performed, and no nerve root injury or dural tear occurred. The average operation time was 138 ± 33 min, intraoperative blood loss was 126 ± 50 mL, the duration from surgery to getting out of bed was 46 ± 8 h, and hospitalization cost was 1.6 ± 0.2 ten thousand yuan. All of the 58 patients were followed up for 7-31 months, with an average of 14.6 months. The postoperative VAS scores and ODI score were significantly improved compared with preoperative data (P < 0.05). The evaluation of the MacNab score was excellent in 41 patients, good in 15, and fair in 2, suggesting an effective rate of 96.6%. The intervertebral height had reduced 0.2 ± 1.2 mm by the last follow-up, and there were 55 Grade I and II cases based on the Bridwell evaluation criterion. The fusion rate was 94.8%, and no screw breakage and loosening occurred. The scores of physical pain, general health, social, and emotional functioning were significantly increased at the last follow-up. CONCLUSION: Minimally invasive transforaminal lumbar interbody fusion combined with posterolateral fusion and unilateral fixation provide a new choice for degenerative lumbar disease, and the short-term clinical outcome is satisfactory.
