Effects of LncRNA GAS6-AS1 on proliferation,apoptosis,and invasion of laryngeal cancer cells by regulating the miR-708-5p/PDK4 axis

XIE Yang; MA Lin; YAO Zhaoxu; LIU Lin; HE Bing

doi:10.3969/j.issn.1009-0460.2025.08.001

Abstract：Objective To investigate the effects of long non-coding RNA(LncRNA)growth arrest-specific gene 6-antisense RNA 1(GAS6-AS1)on the proliferation,apoptosis,and invasion of laryngeal cancer cells by regulating the microRNA-708-5p(miR-708-5p)/pyruvate dehydrogenase kinase isozyme 4(PDK4)axis.Methods The quantitative real-time(qRT-PCR)was applied to detect the expression of LncRNA GAS6-AS1,miR-708-5p,and PDK4 in laryngeal cancer tissue samples.Laryngeal cancer TU686 cells were divided into siRNA negative control(si-NC group),GAS6-AS1 knockdown alone group(si-GAS6-AS1 group),co-transfection with si-GAS6-AS1 and inhibitor negative control group(si-GAS6-AS1+anti-NC group),and co-transfection with si-GAS6-AS1 and anti-miR-708-5p group(si-GAS6-AS1+anti-miR-708-5p group),and the expression level in TU686 cells of each group was detected.The proliferation,apoptosis,migration and invasion of TU686 cells were detected by plate cloning assay,flow cytometry and Transwell assay,respectively.Protein expression was detected by Western blotting method.The dual-luciferase reporter gene assay was used to verify the interaction of genes respectively.Results The expression levels of LncRNA GAS6-AS1 and PDK4 in laryngeal cancer tissue were obviously increased,while the expression level of miR-708-5p was obviously reduced(P<0.05).The expression of LncRNA GAS6-AS1 and PDK4 mRNA,numbers of cloned cells,migrated cells and invasive cells,PCNA,MMP-9,and PDK4 protein expression in TU686 cells in the si-GAS6-AS1 group were lower than those in the si-NC group,and the levels of miR-708-5p,apoptosis rate,and Bax protein expression in the si-GAS6-AS1 group were higher than those in the si-NC group(P<0.05).Compared with the si-GAS6-AS1 group and si-GAS6-AS1+anti-NC group,the expression of PDK4 mRNA,numbers of cloned cells,migrated cells and invasive cells,PCNA,MMP-9,and PDK4 protein expression increased in the si-GAS6-AS1+anti-miR-708-5p group,while the miR-708-5p expression,apoptosis rate,and Bax protein expression decreased(P<0.05).LncRNA GAS6-AS1 targeted and negatively regulated miR-708-5p expression,while miR-708-5p targeted and negatively regulated PDK4 expression.Conclusion Inhibition of LncRNA GAS6-AS1 may downregulate PDK4 expression by targeting miR-708-5p,thereby inhibiting TU686 cell proliferation and invasion,and promoting apoptosis.