摘要
目的:探讨Tob1对肾上腺皮质癌细胞自噬的作用及其影响。方法:将体外培养的SW-13细胞分为对照组、空载质粒转染组以及pcDNA3.1-Tob1质粒转染组。实时荧光定量PCR检测各组细胞中Tob1的mRNA表达。Western blot检测Tob1以及自噬相关蛋白LC3-Ⅰ、LC3-Ⅱ、p62和Beclin 1的表达。LC3免疫荧光检测细胞自噬水平。随后在转染pcDNA3.1-Tob1质粒的SW-13细胞中加入自噬抑制剂3-MA处理,CCK-8检测细胞增殖,Tunel检测细胞凋亡。结果:pcDNA3.1-Tob1质粒转染显著上调SW-13细胞中Tob1的mRNA和蛋白表达,提升LC3-Ⅱ/LC3-Ⅰ和Beclin 1水平,降低p62蛋白表达,并显著提升细胞自噬水平(P<0.05)。与对照组相比,pcDNA3.1-Tob1组SW-13细胞增殖降低,细胞凋亡增加;与pcDNA3.1-Tob1组相比,pcDNA3.1-Tob1+3-MA组细胞增殖提升,细胞凋亡降低(P<0.05)。结论:Tob1能够通过激活细胞自噬降低人皮质癌细胞SW-13增殖并促进其凋亡。
Objective:To study the effect of Tob1 on autophagy of adrenocortical carcinoma cells.Methods:The cultured SW-13 cells were divided into control group,empty plasmid transfection group and pcDNA3.1-Tob1 plasmid transfection group.Real-time quantitative PCR was used to detect Tob1 mRNA expression in each group.Protein expression of Tob1,LC3-Ⅰ,LC3-Ⅱ,p62 and Beclin 1 were measured by Western blot.LC3 immunofluorescence was used to detect cell autophagy level.Subsequently,SW-13 cells transfected with pcDNA3.1-Tob1 were treated with autophagy inhibitor 3-MA.CCK-8 was used to detect cell proliferation and Tunel assay was used to detect cell apoptosis.Results:pcDNA3.1-Tob1 transfection significantly up-regulated the mRNA and protein expression of Tob1,elevated LC3-Ⅱ/LC3-Ⅰand Beclin 1 protein level,decreased p62 expression,and significantly promoted the autophagy level in SW-13 cells(P<0.05).Compared with the control group,SW-13 cell proliferation was decreased and apoptosis was increased in pcDNA3.1-Tob1 group.Compared with pcDNA3.1-Tob1 group,cell proliferation was increased and cell apoptosis was decreased in pcDNA3.1-Tob1+3-MA group(P<0.05).Conclusion:Tob1 can inhibit the proliferation of human cortical adenocarcinoma cell SW-13 and promote its apoptosis by activating autophagy.
作者
李小亮
伦瑞花
LI Xiaoliang;LUN Ruihua(The Second People’s Hospital of Jiaozuo City,He’nan Province 454001;不详)
出处
《医学理论与实践》
2023年第24期4160-4162,4175,共4页
The Journal of Medical Theory and Practice
关键词
肾上腺皮质癌
Tob1
自噬
细胞增殖
细胞凋亡
Adrenocortical carcinoma
Tob1
Autophagy
Cell proliferation
Cell apoptosis
