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Effects of LncRNA H19 on Proliferation Invasion and Radiotherapy Sensitivity of Glioma T98G Cells Through MiR-370-3p/RAF1 Axis and the Mechanism |
XU Hua, et al |
Radiation Therapy Center, Xi'an International Medical Center Hospital, Shaanxi Xi'an 710100, China |
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Abstract Objective: To analyze the effects of lncRNA H19 on proliferation, invasion and radiotherapy sensitivity of brain glioma T98G cells and its mechanism. Methods: The differential expressions of lncRNA H19, miR-370-3p and RAF1 in glioma tissues and adjacent tissues were detected by real-time fluorescence quantitative PCR. The expression levels of lncRNA H19, miR-370-3p and RAF1 were detected after T98G cells were irradiated with X-ray (0, 2, 4, 6 Gy). T98G cells were transfected with H19 siRNA, miR-370-3p inhibitor and RAF1 siRNA plasmid, respectively, after 6 Gy X-ray irradiation, the effects of down-regulation of H19, miR-370-3p and RAF1 on the proliferation, invasion and apoptosis of T98G cells were detected by CCK-8, Transwell and flow cytometry, respectively. The target and correlation between lncRNA H19 and miR-370-3p, miR-370-3p and RAF1 were analyzed by miRcode, TargetScan and dual luciferase reporter gene assay. The effects of upregulated lncRNA H19 on the proliferation, invasion and radiotherapy sensitivity of T98G cells via miR-370-3p were examined. RT-qPCR and Western blot were used to detect the effect of lncRNA H19 on the expression of RAF1 via miR-370-3p. Results: The expressions of lncRNA H19 and RAF1 in glioma tissues were up-regulated (P<0.01), and the expression of miR-370-3p was down-regulated (P<0.01). The expressions of lncRNA H19 and RAF1 decreased after radiation in a dose dependent manner. The expression of miR-370-3p increased after radiation in a dose dependent manner. Downregulation of lncRNA H19 or RAF1 inhibited proliferation and invasion of T98G cells and enhanced radiotherapy sensitivity. LncRNA H19 targeted miR-370-3p and miR-370-3p targeted RAF1. Downregulation of miR-370-3p promoted the proliferation and invasion of T98G cells, and reduced the radiotherapy sensitivity. The up-regulation of lncRNA H19 promoted the proliferation and invasion of T98G cells through miR-370-3p, and reduced the radiotherapy sensitivity. Conclusion: LncRNA H19 regulated the proliferation, invasion and radiotherapy sensitivity of T98G cells through miR-370-3p/RAF1 axis.
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