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作 者:张双美[1] 王斌[1] 陈怡[1] 张小火 施雪霏 ZHANG Shuang-mei;WANG Bin;CHEN Yi(Department of Respiratory Medicine, Huzhou Central Hospital of Zhejiang University y Huzhou, Zhejiang 313000, China)
机构地区:[1]浙江大学湖州医院呼吸内科,浙江湖州313000
出 处:《中华全科医学》2019年第6期897-901,共5页Chinese Journal of General Practice
基 金:国家自然科学基金项目(81602003)
摘 要:目的通过上调或者下调PVT1的表达,分析探究长链非编码RNA PVT1在非小细胞肺癌(NSCLC)细胞增殖和迁移能力中的作用及机制,并在此基础上运用RNA-Seq分析PVT1的靶向基因,说明其作用机理。方法①通过TCGA数据库(The Cancer Genome Atlas)分析NSCLC细胞中PVT1的表达与NSCLC患者生存时间之间的关系;②探究PVT1上调或下调对NSCLC细胞增殖及迁移能力的影响;③运用RNA-seq技术探究PVT1的下游靶基因,然后运用siRNA沉默该基因,探究此基因对NSCLC细胞增殖和迁移能力。结果①PVT1的高表达与肺癌患者存活时间减少有显著关系(P<0.001);②PVT1高表达可以显著促进肺癌患者肿块体积的增大,并且与肿瘤分期和淋巴结转移程度密切相关(均P<0.05);③过表达PVT1可以促进NSCLC细胞的增殖及迁移;④用Si-RNA下调PVT1可以抑制NSCLC细胞的增殖及迁移;⑤PVT1下调会使LATS2的表达水平显著升高。结论 PVT1通过表观调控LATS2促进NSCLC细胞增殖和迁移能力。Objective To investigate the role and mechanism of lncRNA plasmacytoma variant translocation 1(PVT1) in the proliferation and migration of non-small cell lung cancer(NSCLC) cells by up-regulating or down-regulating the expression of PVT1, and then analyze the target gene of PVT1 by using RNA-Seq for its action mechanism. Methods ① The relationship between the expression of PVT1 in NSCLC cells and the survival time of NSCLC patients through studying TCGA database(The Cancer Genome Atlas);② Using Ki67 immunofluorescence assay and cell colony formation assay to investigate the effect of up-regulation or down-regulation of PVT1 on the proliferation and migration of NSCLC cells;③ Using RNA-seq technology to explore the downstream target genes of PVT1, and then use siRNA to silence the gene to explore the ability of this gene to proliferate and invade NSCLC cells. Results ① The high level expression of PVT1 was significantly associated with the decrease of survival time of lung cancer patients(P<0.001).②The high level expression of PVT1 significantly promoted the increase of tumor volume in lung cancer patients, and was closely related to tumor stage and lymph node metastasis(all P<0.05);③ The overexpression of PVT1 promoted the proliferation and migration of NSCLC cells;④Down-regulation of PVT1 by Si-RNA inhibited the proliferation and migration of NSCLC cells;⑤Down-regulation of PVT1 significantly increased the expression level of LATS2. Conclusion PVT1 can promote the proliferation and migration of NSCLC cells by apparently regulating LATS2.
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