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@#[Abstract] Objective: To study the expression of miRNA-95 in osteosarcoma tissues and cell lines, as well as to reveal its effect on proliferation, apoptosis, cell cycle and invasion ability of osteosarcoma cells. Methods: Real-time fluorescent quantitative PCR was used to detect the expression of miRNA-95 in 15 pairs of osteosarcoma tissues and their adjacent normal tissues (specimens were collected from patients underwent surgery in Qingdao Haici Medical Group from January 2015 to January 2018), osteosarcoma cell lines (MG-63, U2OS, 143B and HOS) and normal human osteoblast hFOB1.19 cell line. miRNA-95 mimics and miRNA-95 inhibitors were respectively transfected into MG-63 cells by Lipofectamine 2000, and miRNA-NC group was set up as control group. CCK-8 method was used to detect the changes in cell proliferation, Flow cytometry was used to detect the changes in cell cycle and apoptosis, Transwell method was used to detect the changes in cell invasion ability, and Dual luciferase enzyme activity assay was used to detect and validate the target gene of miRNA-95 in osteosarcoma cells. Results: The expression level of miRNA-95 in human osteosarcoma tissues and cell lines (MG-63, U2OS, 143B and HOS) was significantly higher than that in adjacent tissues and normal human osteoblast hFOB1.19 cell line (all P<0.01), with the highest expression in MG-63 cells (P<0.01). Compared with the miRNA-NC group, the proliferation and invasion abilities of MG-63 cells in miRNA-95 mimics group increased significantly, while the apoptosis rate decreased significantly (all P<0.01). However, the proliferation and invasion activities of MG-63 cells in miRNA-95 inhibitor group decreased significantly, while the apoptosis rate increased significantly, and the cell cycle was obviously blocked (all P<0.01). miRNA-95 played a role in targeting the gene of epithelialmembraneprotein1 (EMP-1) in human osteosarcoma MG-63 cells. Conclusion: miRNA-95 is highly expressed in human osteosarcoma tissues and cells; inhibitor of miRNA-95 expression can promote apoptosis and inhibit proliferation, cell cycle and invasion of osteosarcoma cells, which may be related with targeting EMP-1 gene.
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Objective To detect the expression of miRNA-95 in cervical cancer tissues and cell lines with different radiosensitivity and study the effect of its regulation on radiosensitivity of cervical cancer cells. Methods Real-time PCR was used to detect the expression of miRNA-95 in cervical cancer tissues of 20 patients with radiosensitivity, 20 patients with radiation tolerance, radioresistant cervical cancer cell lines (HeLa, SiHa), and radiosensitive cervical cancer cell lines (Me180). MiRNA-95 mimics (miRNA-95 mimics group ) and miRNA-95 inhibition ( miRNA-95 inhibition group ) were transfected into radioresistant HeLa and SiHa cells by liposome 2000, miRNA-NC was set as control group. CCk-8 assay was used to detect the proliferation of cervical cancer cells irradiated with 60Co γ-rays at 0,2,4,6,8, 10 Gy. After 4 Gy irradiation, cell clonal formation ability was detected by plate monoclonal assay, and cell apoptosis was detected by flow cytometry. Dual luciferase activity assay was used to detect the target gene of miRNA-95 in cervical cancer cells. Nude mice were used to detect the changes of tumor formation ability. Results The expression of miRNA-95 in cervical cancer tissues of patients with radiotherapy tolerance was significantly higher than that of patients with radiotherapy sensitivity ( t =12. 279, P <0. 05 ) . The expressions of miRNA-95 in HeLa and SiHa cells were significantly higher those that of Me180 cells (t = 5.162, 7.114, P < 0.05). When the cells were treated with miRNA-95 inhibition, the expression of miRNA-95 in HeLa and SiHa cell lines was significantly lower than that of microRNA-NC group (t =5. 162, 7. 114, P < 0. 05), the cell proliferation rate decreased significantly ( t =8. 273, 11. 354, 13. 489, 15. 396 and 6. 197, 9. 185, 10. 994, 12. 442, P<0. 05), the cell monoclonal formation rate decreased significantly (t=8. 378, 7. 931, P<0. 05), and the apoptosis rate increased significantly (t=10. 265, 8. 386, P<0. 05). The tumorigenic weight of nude mice in the miRNA95 inhibition group was significantly decreased (t=8. 881, 10. 037, P<0. 05). Conclusions The miRNA-95 had low levels in both radiosensitive cervical cancer tissues and cells. Inhibiting the expression of microRNA-95 can significantly improve the radiosensitivity of cervical cancer cells by targeting SGPP1 gene.