Role of ST8SIA3 in temozolomide resistance of glioma cells
Deng Zhitong
Zhao Zhaohui
Cai Yong
Zhong Xingming
Wang Yiqi
Yang Jianguo
Fei Zhenhai
Zhang Lei
Gu Hua
Yang Tao
Abstract：Objective To investigate the influence of ST8 α-N-acetylneuraminidase α-2,8-sialyltransferase 3(ST8SIA3)on the development of temozolomide(TMZ)resistance in glioma cells.Methods The glioblastoma U87-MG cell lines were subjected to lentiviral transfection and divided into three groups:control group,downregulation group(reduced ST8SIA3 expression)and overexpression group(increased ST8SIA3 expression).The relative expressions of ST8SIA3 in these groups were detected using qRT-PCR,while A2B5 and O6-methylguanine-DNA methyltransferase(MGMT)expressions were analyzed via Western-blot.After culturing the three groups with varying concentrations of TMZ(0µmol/L,5µmol/L,10µmol/L,20µmol/L,50µmol/L,100µmol/L,200µmol/L)for 48 hours,the TMZ concentration-cell survival curves were plotted using CCK-8 assays.The effects of ST8SIA3 in downregulation group and overexpression group on the self-renewal capacity of glioma stem cells(GSC)and stem cell markers(CD133,SOX2)were observed through single-cell sphere formation experiments.The relationship between ST8SIA3 expression and patient prognosis was analyzed using clinical data from The Cancer Genome Atlas(TCGA)and Chinese Glioma Genome Atlas(CGGA)databases.Survival analysis and the recording of the onset of cachexia were conducted in nude mouse tumor models,and immunohistochemical methods were used to detect MGMT and A2B5 expressions in orthotopic transplanted tumor samples from nude mice.Results The relative expressions of A2B5 and MGMT increased in the overexpression group.CCK-8 assay results showed that the LC50(lethal concentration 50%)of TMZ gradually increased with increase of ST8SIA3 expression.The single-cell sphere formation experiments demonstrated that the number and size of spheres gradually increased with increase of ST8SIA3 expression.Results from the nude mouse orthotopic tumor models indicated that overexpression of ST8SIA3 significantly shortened the survival period of nude mice,and expressions of Ki-67,A2B5 and MGMT increased with increase of ST8SIA3 expression.Conclusions ST8SIA3 promotes the sphere-forming ability of U87-MG cells by synthesizing the stem cell marker A2B5,thereby conferring GSC resistance to TMZ.
Keywords：gliomaglioma stem cellstemozolomidedrug resistanceST8SIA3A2B5
Publication Date：2024-07-20
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 424-430 )