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LncRNA MIR100HG Regulation of the miR-142-5p / SOX 5 axis on the proliferation and invasion ability of oral squamous carcinoma cells |
WEI chao1, DONG Zihan2, AN ning1, YANG Qian1, MA Yongping1 |
1. Department of Stomatology, Baoding Second Hospital, Baoding 071000, China; 2. Department of Human Anatomy and Embryology, Hebei North University, Zhangjiakou 075000, China |
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Abstract Objective To investigate the effect of the long non-coding RNA mir-100-let-7a-2-ir-125b-1 cluster host gene (LncRNA MIR100HG) regulating miR-14-2-5p / SRY related high mobility box protein 5(SOX 5) axis on the proliferation and invasion capacity of oral squamous cell carcinoma (OSCC) cells. Methods Quantitative real-time PCR and immunoblot were performed to detect the expression of LncRNA MIR100HG, miR-142-5p and SOX 5 in CAL 27, SAS, and SCC-25 in vitro cultured human oral epithelial cells and human OSCC-derived cells. When CAL 27 cells were cultured in vitro, They were randomly divided into control group, LncRNA MIR100HG knockdown group (transfection with LncRNA MIR100HG siRNA plasmid), miR-142-5p mimics group (transfected with miR-142-5p mimics), cotransfection negative control group (transfection with empty plasmid and miR-142-5p negative control), LncRNA MIR100HG knockdown + miR-142-5p inhibitor group (transfected with LncRNA MIR100HG The siRNA plasmid and miR-142-5p inhibitor), After the group-transfection, The expression of LncRNA MIR100HG, miR-142-5p and SOX 5 were analyzed by real-time PCR and immunoblotting; Using the method 3- (4, 5-dimethylthiazole-2) -2, 5-diphenyl tetrazolium bromine salt (MTT) method, 5-ethylene-2-2 ′ -deoxyuridine (Edu) staining and colony formation assay to detect cell proliferation in each group; CAL 27 cell invasion in each group was detected by Transwell invasion assay; Immunoblotting was used to detect protein expression associated with epithelial stromal transformation in each group of CAL 27 cells. OSCC tumor model was constructed in the area near the right armpit of the back of nude mice. The tumor volume and weight of nude mice were detected after 3 weeks of feeding. A dual luciferase reporter assay was used to detect the targeted regulation of miR-142-5p and SOX 5 by miR-142-5p. Results Compared with human oral epithelial cells, LncRNA MIR100HG, SOX 5 protein and mRNA were increased in CAL 27, SAS, SCC-25 and miR-142-5p decreased. In LncRNA MIR100HG knockdown, SOX 5 protein and mRNA expression, cell viability, proliferation rate, colony formation ratio, tumor volume and weight, invasion number, N-cadherin protein expression, miR-142-5p expression than control group. Downregulation of miR-142-5p can weaken the effect of LncRNA MIR100HG knockdown on various indicators of CAL27 cells. Targeted downregulation of miR-142-5p by LncRNA MIR100HG and miR-142-5p and SOX 5 in CAL 27 cells. Conclusion Knockdown of LncRNA MIR100HG can inhibit OSCC cells proliferation, epithelial stromal transformation and invasion by regulating the miR-142-5p/SOX5 axis, and can delay tumor growth in vivo.
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Received: 08 December 2022
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