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Study on the Mechanism of miR-216a-3p and miR-128-3p Inhibiting the Phenotypic Deterioration of Oral Squamous Cell Carcinoma Cells by Regulating RGS17 |
WANG Hui |
Jinan Hospital of Traditional Chinese Medicine, Shandong Jinan 250012, China |
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Abstract Objective: To observe the effects of miR-216a-3p and miR-128-3p on the phenotypic expression of OSCC cells,and explore their potential mechanisms. Methods: The expression of miR-216a-3p,miR-128-3p and G protein signal regulator 17 (RGS17) in cancer tissues,adjacent tissues,human normal oral keratinocytes (hNOK) and OSCC cancer cells (SCC-9,SCC-25,HN4) were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-qPCR); mimic NC group (transfected mimic),mimic-miR-216a-3p group (transfected miR-216a-3p mimic),mimic-miR-128-3p group (transfected miR-128-3p mimic),si-NC group (transfected si-NC),si-RGS17 group (transfected si-RGS17),mimic-miR-216a-3p+pcDNA group (co-transfected miR-216a-3p mimic and pcDNA),and mimic-miR-216a-3p+pcDNA-RGS17 group(co-transfected miR-216a-3p mimic and pcDNA-RGS17) were transfected into SCC-9 cells.5-bromo-2-deoxyuracil (EdU) staining and clonogenic assay were used to detect cell proliferation; Transwell test was used to detect the ability of cell migration and invasion; Annexin V / propidium iodide (PI) staining was used to detect the ability of apoptosis; Double luciferase report assay and RNA sedimentation assay were used to verify the targeting relationship; The RGS17 protein was detected by Western blot (WB). Results: Compared with adjacent tissues or hNOK cells,the expression of miR-216a-3p and miR-128-3p in OSCC tissues (stage Ⅰ,Ⅱ,Ⅲ) and cancer cells (SCC-9,SCC-25,HN4) was significantly decreased,and the expression of RGS17 was significantly increased (P<0.05).Compared with the mimic-NC group,the positive rate of EdU,the rate of clone formation,the number of migrating and invasive cells in the mimic-miR-216a-3p and mimic-miR-128-3p groups were significantly lower,and the apoptosis rate was significantly higher (P<0.05).The cells in si-RGS17 group had similar changes to those in mimic-miR-216a-3p and mimic-miR-128-3p cells.miR-216a-3p and miR-128-3p jointly targeted and negatively regulated RGS17.Overexpression of RGS17 significantly attenuated the proliferation,migration-invasion inhibition and apoptosis-promoting effects of miR-216a-3p and miR-128-3p on SCC-9 cells. Conclusion: MiR-216a-3p and miR-128-3p could inhibit the proliferation,migration and invasion of OSCC cells and promote apoptosis,which may be related to targeting RGS17.
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