摘要
目的探究LINC01354调控脑胶质瘤细胞增殖、凋亡及焦亡的功能及机制。方法实时荧光定量聚合酶链反应(qRT-PCR)检测人小胶质细胞HMC3和脑胶质瘤细胞U251中LINC01354相对表达量。双荧光素酶报告基因实验验证LINC01354和miR-214-3p的靶向关系。人脑胶质瘤细胞U251分为空白组、sh-NC组、sh-LINC01354组、miR-214-3p mimic组、miR-NC组、sh-LINC01354+miR-214-3p inhibitor组和sh-LINC01354+inhibitor NC组。MTT法和流式细胞术检测细胞增殖和凋亡。Western blot检测细胞焦亡相关蛋白NOD样受体家族吡喃结构域包含蛋白3(NLRP3)、剪切的半胱天冬氨酸蛋白酶-1(Cleaved Caspase-1)和剪切的N-末端消皮素D(Cleaved N-terminal GSDMD)蛋白表达。结果人脑胶质瘤细胞U251中LINC01354表达高于人小胶质细胞HMC3(P<0.001),双荧光素酶报告基因结果显示miR-214-3p为LINC01354靶基因。MTT和流式细胞术检测结果显示,sh-LINC01354组U251细胞增殖能力低于sh-NC组(P<0.05),细胞凋亡高于sh-NC组(P<0.001),NLRP3、Cleaved Caspase-1和Cleaved N-terminal GSDMD蛋白表达均高于sh-NC组(P<0.01)。miR-214-3p mimic组U251细胞增殖能力均低于miR-NC组(P<0.05),细胞凋亡高于miR-NC组(P<0.001),NLRP3、Cleaved Caspase-1和Cleaved N-terminal GSDMD蛋白表达均高于miR-NC组(P<0.001)。sh-LINC01354+miR-214-3p inhibitor组U251细胞增殖高于sh-LINC01354+inhibitor NC组(P<0.05),细胞凋亡低于sh-LINC01354+inhibitor NC组(P<0.001)。结论LINC01354靶向抑制miR-214-3p影响U251细胞的增殖、凋亡及焦亡。
Objective To investigate the function and mechanism of LINC01354 in regulating the proliferation,apoptosis and pyroptosis of glioma cells.Methods qRT-PCR was used to detect the relative expression levels of LINC01354 in HMC3 and U251 Cells.The dual luciferase reporter gene experiment validated the targeting relationship between LINC01354 and miR-214-3p.Human glioma cells U251 were divided into blank group,sh-NC group,sh-LINC01354 group,miR-214-3p mimic group,miR-NC group,sh-LINC01354+miR-214-3p inhibitor group and sh-LINC01354+inhibitor NC group.MTT assay and flow cytometry were used to detect cell proliferation and apoptosis.Western blot was used to detect the expres-sion of NLRP3,Cleaved Caspase-1 and Cleaved N-terminal GSDMD protein.Results The relative expres-sion levels of LINC01354 in U251 cells were significantly higher than those in HMC3 cells(P<0.001).The dual luciferase reporter gene results showed that miR-214-3p was the target gene of LINC01354.The MTT and flow cytometry detection results showed that the proliferation ability of U251 cells in the sh-LINC01354 group was significantly lower than that in the sh-NC group(P<0.05),the apoptosis was sig-nificantly higher than that in the sh-NC group(P<0.001),the protein expression of NLRP3,Cleaved Caspase-1 and Cleaved N-terminal GSDMD were significantly higher than that in the sh-NC group(P<0.01).The proliferation ability of U251 cells in the miR-214-3p mimic group was significantly lower than that in the miR-NC group(P<0.05),cell apoptosis was significantly higher than that in the miR-NC group(P<0.001),the protein expression of NLRP3,Cleaved Caspase-1 and Cleaved N-terminal GSDMD were significantly higher than that in the miR-NC group(P<0.001).The proliferation of U251 cells in the sh-LINC01354+miR-214-3p inhibitor group was significantly higher than that in sh-LINC01354+in-hibitor NC group(P<0.05),and the apoptosis was lower than that in sh-LINC01354+inhibitor NC group(P<0.001).Conclusion LINC01354 inhibites miR-214-3p to affect the proliferation,apoptosis and pyroptosis of U251 cells.
作者
徐敬轩
杨操
葛畅
张文杰
栾新平
XU Jingxuan;YANG Cao;GE Chang;ZHANG Wenjie;LUAN Xinping(The Second Affiliated Hospital of Xinjiang Medical University,Urumqi 830028,China)
出处
《新疆医科大学学报》
2025年第9期1226-1233,共8页
Journal of Xinjiang Medical University
基金
新疆维吾尔自治区区域协同创新专项(新疆维吾尔自治区科技支疆项目)(2022E02060)。
