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The effects of aberrant expression of LncRNA DGCR5/miR‐873‐5p/TUSC3 in lung cancer cell progression

机译:LncRNA DGCR5 / miR-873-5p / TUSC3异常表达对肺癌细胞进展的影响

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摘要

Lung cancer is the most common cause of cancer‐related mortality worldwide, and nonsmall cell lung cancer (NSCLC) accounts for 80% of all pulmonary carcinomas. Recently, long noncoding RNAs (lncRNAs) have been paid attention for exploring treatment of various diseases. Upregulation of DiGeorge syndrome critical region gene 5 (DGCR5) predicts better lung squamous cell carcinoma prognosis; therefore, we explore the role of DGCR5 in lung cancer in our present study. Consecutive patients with LC were treated in our hospital between January 2015 and January 2016. qRT‐PCR demonstrated that DGCR5 was significantly lower in neoplastic tissues than in non‐neoplastic tissues. For in vitro experiments, cell growth, migration, and invasion were significantly lower in A549 cells transfected with pcDNA3.1‐DGCR5 than pcDNA3.1, which were verified by 5‐diphenyltetrazolium bromide (MTT) assay, scratch test, and transwell assay, respectively, with no significant induction on cell apoptosis that was demonstrated by flow cytometry (FCM) assay. Bioinformatics analysis predicted that 3’ untranslated region (UTR) of tumor suppressor candidate 3 (TUSC3, 49‐55 bp) and style="fixed-case">DGCR5 (801‐807 bp) shared a common hsa‐miR‐873‐5p binding site, and the direct interaction between style="fixed-case">DGCR5 and hsa‐miR‐873‐5p or hsa‐miR‐873‐5p and style="fixed-case">TUSC3 was verified by dual‐luciferase reporter assay. style="fixed-case">qRT‐ style="fixed-case">PCR demonstrated that hsa‐miR‐873‐5p was dramatically higher and style="fixed-case">TUSC3 was significantly lower in neoplastic tissues than in non‐neoplastic tissues. style="fixed-case">DGCR5 decreased the protein level of style="fixed-case">TUSC3 by miR‐873‐5p which was demonstrated by Western blot and immunofluorescence. The role of style="fixed-case">DGCR5 in tumorigenesis in vivo was consistent with in vitro assays, Ki‐67‐positive cell number (exhibited by immunohistochemical staining), tumor size, and tumor weight of A549‐ style="fixed-case">DGCR5 group were significantly lower in comparison with A549‐control group.
机译:肺癌是世界范围内与癌症相关的死亡率的最常见原因,非小细胞肺癌(NSCLC)占所有肺癌的80%。近年来,长的非编码RNA(lncRNA)已被关注用于探索各种疾病的治疗。 DiGeorge综合征关键区域基因5(DGCR5)的上调预示着肺鳞癌的预后更好。因此,在本研究中,我们探讨了DGCR5在肺癌中的作用。连续LC患者在2015年1月至2016年1月间在我院接受治疗。qRT-PCR表明,肿瘤组织中的DGCR5明显低于非肿瘤组织中的DGCR5。在体外实验中,pcDNA3.1-DGCR5转染的A549细胞的细胞生长,迁移和侵袭比pcDNA3.1显着降低,这已通过5-二苯基溴化四氮唑(MTT)分析,刮擦试验和transwell分析进行了验证,分别对流式细胞术(FCM)分析所证实的细胞凋亡无明显诱导作用。生物信息学分析预测,抑癌候选基因3(TUSC3,49-55bp)和 style =“ fixed-case”> DGCR 5(801-807bp)的3'非翻译区(UTR)共有一个共同点hsa‐miR‐873‐5p结合位点,以及 style =“ fixed-case”> DGCR 5与hsa‐miR‐873‐5p或hsa‐miR‐873‐5p和之间的直接相互作用通过双重荧光素酶报告基因分析验证了style =“ fixed-case”> TUSC 3。 style =“ fixed-case”> qRT - style =“ fixed-case”> PCR 表明hsa‐miR‐873-5p显着提高,而 style =“ fixed肿瘤组织中的-case“> TUSC 3显着低于非肿瘤组织。 Western blot和Western blot证实 style =“ fixed-case”> DGCR 5降低了miR‐873-5p的 style =“ fixed-case”> TUSC 3的蛋白质水平。免疫荧光。 style =“ fixed-case”> DGCR 5在体内肿瘤发生中的作用与体外测定,Ki-67阳性细胞数(免疫组织化学染色显示),肿瘤大小和肿瘤重量一致与A549对照组相比,A549- style =“ fixed-case”> DGCR 5组的Aβ显着降低。

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