LncRNA-CASC2竞争性结合miR-193a-5p调控GSDMD焦亡通路逆转非小细胞肺癌顺铂耐药的分子机制
Molecular mechanisms of LncRNA-CASC2 competitively binding miR-193a-5p regulating GSDMD scorch death pathway reversing Cisplatin re-sistance in non-small cell lung cancer
罗世程 1肖冬平 1黄秀玲 1左炜 1胡向东 1邓志辉1
作者信息
- 1. 南昌市第一医院全科医学科,江西南昌 330006
- 折叠
摘要
目的 探讨LncRNA-CASC2竞争性结合miR-193a-5p调控GSDMD焦亡通路逆转非小细胞肺癌顺铂耐药的分子机制.方法 选用非小细胞肺癌(NSCLC)细胞系A549进行研究,建立耐药株A549/DDP,通过CCK8检测细胞对顺铂的敏感性验证耐药细胞株是否成功建立.利用PCR检测CASC2在A549和A549/DDP细胞中的表达情况以及对GSDMD2、miR-193a-5p表达的影响.通过生物信息学软件预测LncRNA-CASC2与miR-193a-5p、miR-193a-5p与GSDMD结合位点,利用荧光素酶报告基因分析CASC2与miR-193a-5p、miR-193a-5p与GSDMD靶向关系,Western blot检测LncRNA-CASC2竞争性结合miR-193a-5p调控GSDMD焦亡通路的蛋白表达变化.结果 A549/DDP细胞的半抑制浓度值是A549细胞的10倍以上,表明已成功构建耐药细胞株A549/DDP.A549/DDP中CASC2表达较细胞A549降低(P＜0.05).CASC2 mimics可抑制细胞中miR-193a-5p表达,同时GSDMD 表达上调(P＜0.05).CASC2 与 miR-193a-5p、miR-193a-5p 与 GSDMD 具有明显靶向关系(P＜0.05).结论 非小细胞肺癌细胞中LncRNA-CASC2、miR-193a-5p低表达与GSDMD的高表达,与非小细胞肺癌化疗耐药性密切相关,LncRNA-CASC2与miR-193a-5p相互作用调控GSDMD,促进肿瘤细胞耐药.
Abstract
Objective To explore the molecular mechanism of the competitive binding of LncRNA-CASC2 to miR-193a-5p in regulating GSDMD focal death pathway to reverse cisplatin resistance in non-small cell lung cancer.Methods Non-small cell lung cancer(NSCLC)cell line A549 was selected to establish a drug-resistant strain A549/DDP,and the sensitivity of the cell to cisplatin was detected by CCK8 to verify whether the drug-resistant cell line was success-fully established.The expression of CASC2 in A549 and A549/DDP cells and its effect on the expression of GSDMD2 and miR-193a-5p were detected by PCR.The binding sites of LncRNA-CASC2 and miR-193a-5p,miR-193a-5p and GSD-MD were predicted by bioinformatics software.The target relationship between CASC2 and miR-193a-5p and miR-193a-5p and GSDMD was analyzed by luciferase reporter gene,and the competitive binding of LncRNA-CASC2 to miR-193a-5p was detected by Western blot.Results The semi-inhibitory concentration of A549/DDP cells was more than 10 times that of A549 cells,indicating that the drug-resistant cell line A549/DDP had been successfully constructed.The expression level of CASC2 in A549/DDP was lower than that in A549(P＜0.05).CASC2 mimics inhibited the expression of miR-193a-5p in cells,while GSDMD expression was up-regulated(P＜0.05).There was a significant targeting relationship between CASC2 and miR-193a-5p,and between miR-193a-5p and GSDMD(P＜0.05).Conclusion The low expression of LncRNA-CASC2 and miR-193a-5p in NSCLC cells and the high expression of GSDMD are closely related to the chemotherapy resistance of NSCLC.The interaction of LncRNA-CASC2 and miR-193a-5p regulates GSDMD and promotes the drug resistance of tumor cells.
关键词
LncRNA-CASC2/miR-193a-5p/GSDMD/耐药Key words
LncRNA-CASC2/miR-193a-5p/GSDMD/Drug resistance引用本文复制引用
基金项目
江西省卫生健康委科技项目(202211574)
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据