Effects of celastrol on the proliferation,migration,and invasion of oral squamous cell carcinoma cells by regulating miR-216a-3p/FN1

LIU Ya-juan; DUAN Feng; WANG Xiao-yu; WANG Yi-yuan

doi:10.19749/j.cn.cjgd.1672-2973.2024.06.002

Abstract：Objective To investigate the effects of celastrol (CEL) on the proliferation,migration and invasion of oral squamous cell carcinoma (OSCC) cells by regulating microRNA-216a-3p (miR-216a-3p)/fibronectin 1 (FN1). Methods MTT was used to detect the effects of different concentrations of CEL on the survival rate of human OSCC cells (SCC-25) in cultured vitro,and to screen out the best working concentration of CEL. SCC-25 cells were randomly divided into Control group,L-CEL group,H-CEL group,H-CEL+anti-NC group and H-CEL+anti-miR-216a-3p group. The mRNA expression of miR-216a-3p and FN1 mRNA were detected by qRT-PCR. Dual luciferase assay was used to detect the relationship between miR-216a-3p and FN1. MTT and colony formation assays were used to detect cell proliferation. Cell migration was detected by scratch test. Transwell assay was used to detect cell invasion. Cell apoptosis was detected by flow cytometry. Western blotting was used to detect the protein expression in cells. Results Compared with the Control group,the L-CEL group and H-CEL group showed increased cell apoptosis rate,miR-216a-3p expression,and E-cadherin expression,as well as decreased cell viability,cloning number,proliferating cell nuclear antigen (PCNA) expression,wound healing rate,invasion number,Vimentin expression,and FN1 expression (P<0.05). Compared with the H-CEL group and H-CEL+anti-NC group,the cell apoptosis rate,miR-216a-3p,and E-cadherin expression were decreased in the H-CEL+anti-miR-216a-3p group,while the cell survival rate,clone number,PCNA,scratch healing rate,invasion number,Vimentin,and FN1 expression were increased (P<0.05). Dual luciferase assay showed that after transfection with WT-FN1,the luciferase activity of the miR-216a-3p mimic group decreased (P<0.05). Conclusion CEL may induce apoptosis of OSCC SCC-25 cells and inhibit proliferation,migration,and invasion of SCC-25 cells by upregulating miR-216a-3p and targeting downregulating FN1 expression.