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LncRNA NALT1在Erastin诱导的结直肠癌细胞铁死亡中的作用及机制研究
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作者 吴金露 姜丽霞 +2 位作者 林艳艳 谢雨萱 叶梦玲 《广西医科大学学报》 2025年第4期543-551,共9页
目的:初步探讨长链非编码RNA(lncRNA)NALT1结合抑癌蛋白p53,促进Erastin介导的人类结直肠癌(CRC)细胞铁死亡的作用及其机制。方法:铁死亡诱导剂Erastin诱导RKO细胞发生铁死亡,使用流式细胞术、细胞计数试剂盒(CCK-8)检测细胞活力,并评... 目的:初步探讨长链非编码RNA(lncRNA)NALT1结合抑癌蛋白p53,促进Erastin介导的人类结直肠癌(CRC)细胞铁死亡的作用及其机制。方法:铁死亡诱导剂Erastin诱导RKO细胞发生铁死亡,使用流式细胞术、细胞计数试剂盒(CCK-8)检测细胞活力,并评估细胞死亡敏感性。同时使用实时定量聚合酶链反应(qPCR)检测细胞内NALT1表达水平。通过慢病毒感染构建NALT1稳定过表达RKO细胞株和对照细胞株,进一步使用流式细胞术和免疫荧光检测NALT1对Erastin诱导的脂质过氧化的影响。铁死亡抑制剂Ferrostatin-1(Fer-1)干预实验用于评估NALT1对Erastin介导的细胞活力变化、脂质过氧化水平及谷胱甘肽(GSH)含量的影响。RNA免疫共沉淀实验检测NALT1与p53蛋白的相互作用。此外,通过流式细胞术和免疫荧光评估过表达NALT1是否能够恢复敲低p53对Erastin诱导的脂质过氧化的抑制作用。结果:Erastin处理后,RKO细胞脂质过氧化水平显著升高,细胞活力下降,且细胞对Erastin的铁死亡敏感性增强,同时NALT1表达显著上调(P<0.05)。NALT1过表达可进一步增强Erastin介导的脂质过氧化和细胞铁死亡(P<0.05)。Fer-1处理显著缓解了Erastin诱导的脂质过氧化和细胞铁死亡,并部分逆转NALT1促进的脂质过氧化反应。在GSH含量检测实验中,Fer-1可在对照组中部分恢复GSH水平;但在NALT1过表达细胞中,该恢复作用不明显。结论:NALT1可能通过结合p53,上调CRC细胞脂质过氧化水平并增强对Erastin的敏感性,促进铁死亡。 展开更多
关键词 lncRNA nalt1 Erastin 结直肠癌 铁死亡 P53 Ferrostatin-1
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Long noncoding RNA NALT1-induced gastric cancer invasion and metastasis via NOTCH signaling pathway 被引量:11
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作者 Hai-Yan Piao Shuai Guo +1 位作者 Yue Wang Jun Zhang 《World Journal of Gastroenterology》 SCIE CAS 2019年第44期6508-6526,共19页
BACKGROUNDLong noncoding RNAs (lncRNAs) are aberrant and play critical roles in gastriccancer (GC) progression and metastasis. Searching for coexpressed lncRNAclusters or representative biomarkers related to malignant... BACKGROUNDLong noncoding RNAs (lncRNAs) are aberrant and play critical roles in gastriccancer (GC) progression and metastasis. Searching for coexpressed lncRNAclusters or representative biomarkers related to malignant phenotypes of GC mayhelp to elucidate the mechanism of tumor development and predict the prognosisof GC.AIMTo investigate the prognostic value of NOTCH1 associated with lncRNA in T cellacute lymphoblastic leukemia 1 (NALT1) in GC and the mechanism of itsinvolvement in GC invasion and metastasis.METHODSRNA sequencing and corresponding clinical data were downloaded from TheCancer Genome Atlas database. The significance module was studied byweighted gene coexpression network analysis. A total of 336 clinical sampleswere included in the study. Gene silencing, reverse transcription polymerasechain reaction, western blotting, scrape motility assay, and Transwell migrationassay were used to assess the function of hub-lncRNAs.RESULTSAt the transcriptome level, 3339 differentially expressed lncRNAs were obtained.weighted gene coexpression network analysis was used to obtain 15 lncRNAclusters and observe their coexpression. Pearson’s correlation showed that blue module was correlated with tumor grade and survival. NALT1 was the hublncRNAof blue module and was an independent risk factor for GC prognosis.NALT1 was overexpressed in GC and its expression was closely related toinvasion and metastasis. The mechanism may involve NALT1 regulation ofNOTCH1, which is associated with lncRNA in T cell acute lymphoblasticleukemia, through cis regulation, thereby affecting the expression of the NOTCHsignaling pathway.CONCLUSIONNALT1 is overexpressed and promotes invasion and metastasis of GC. Themechanism may be related to regulation of NOTCH1 by NALT1 and its effect onNOTCH signaling pathway expression. 展开更多
关键词 GASTRIC CANCER Weighted gene COEXPRESSION network analysis nalt1 NOTCH1 CANCER SURVIVAL
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