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Abstract：
OBJECTIVE: It has been verified that long non-coding RNAs (lncRNAs) play critical roles in the development of human cancers. Increasing evidence indicates that lncRNA human plasmacytoma variant translocation1 (PVT1) was dysregulated in non-small cell lung cancer (NSCLC) which is the leading cause of cancer-related death. However, the precise mechanism underlying the effect of PVT1 remains elusive. Our research focused on the correlation of PVT1 to miR-148 and RAB34 in NSCLC.
METHODS: The quantitative real-time PCR (qRT-PCR) and western blot assay were used to detect gene and protein expression in NSCLC tissues and cells. CCK8, colony formation, transwell and wound healing assays were performed to evaluate the cell function of NSCLC cells. Dual-luciferase activity assay and RNA pull down assays were performed to verify the interaction between miR-148 and its targets. A xenograft test was conducted to detect the impact of RAB34 on tumor development in vitro.
RESULTS: In NSCLC tissues and cells, PVT1 and RAB34 were up-regulated, and miR-148 was down-regulated. Overexpression of PVT1 was capable of promoting NSCLC cell proliferation and migration which could be reversed by miR-148 restoration or RAB34 knock down. Also, our data firstly determined that the down-regulation of RAB34 had inhibitor effects while the up-regulation of RAB34 had promotive effects on tumor growth in vitro and in vivo.
CONCLUSIONS: Those findings indicated that the signal pathway PVT1/miR-148/RAB34 play critical roles in the progression of NSCLC could be proposed in NSCLC as a possible diagnosis or therapeutic targets.
METHODS: The quantitative real-time PCR (qRT-PCR) and western blot assay were used to detect gene and protein expression in NSCLC tissues and cells. CCK8, colony formation, transwell and wound healing assays were performed to evaluate the cell function of NSCLC cells. Dual-luciferase activity assay and RNA pull down assays were performed to verify the interaction between miR-148 and its targets. A xenograft test was conducted to detect the impact of RAB34 on tumor development in vitro.
RESULTS: In NSCLC tissues and cells, PVT1 and RAB34 were up-regulated, and miR-148 was down-regulated. Overexpression of PVT1 was capable of promoting NSCLC cell proliferation and migration which could be reversed by miR-148 restoration or RAB34 knock down. Also, our data firstly determined that the down-regulation of RAB34 had inhibitor effects while the up-regulation of RAB34 had promotive effects on tumor growth in vitro and in vivo.
CONCLUSIONS: Those findings indicated that the signal pathway PVT1/miR-148/RAB34 play critical roles in the progression of NSCLC could be proposed in NSCLC as a possible diagnosis or therapeutic targets.
摘要：
目的：已经证实长链非编码RNA（lncRNAs）在人类癌症的发展中起着关键作用。越来越多的证据表明，lncRNA人浆细胞瘤变体易位1（PVT1）在非小细胞肺癌（NSCLC）中失调，这是癌症相关死亡的主要原因。然而,PVT1效应的确切机制仍然难以捉摸。我们的研究集中在NSCLC中PVT1与miR-148和RAB34的相关性。
方法：采用实时荧光定量PCR(qRT-PCR)和免疫印迹法检测NSCLC组织和细胞中基因和蛋白的表达。CCK8，菌落形成，进行了transwell和伤口愈合试验以评估NSCLC细胞的细胞功能。进行双荧光素酶活性测定和RNA下拉测定以验证miR-148与其靶标之间的相互作用。进行异种移植试验以检测RAB34对体外肿瘤发展的影响。
结果：在NSCLC组织和细胞中，PVT1和RAB34上调，miR-148下调。PVT1的过表达能够促进NSCLC细胞增殖和迁移，这可以通过miR-148恢复或RAB34敲低来逆转。此外，我们的数据首先确定RAB34的下调具有抑制作用,而RAB34的上调在体外和体内对肿瘤生长具有促进作用.
结论：这些结果表明，信号通路PVT1/miR-148/RAB34在NSCLC的进展中起关键作用，可作为NSCLC的可能诊断或治疗靶点。
方法：采用实时荧光定量PCR(qRT-PCR)和免疫印迹法检测NSCLC组织和细胞中基因和蛋白的表达。CCK8，菌落形成，进行了transwell和伤口愈合试验以评估NSCLC细胞的细胞功能。进行双荧光素酶活性测定和RNA下拉测定以验证miR-148与其靶标之间的相互作用。进行异种移植试验以检测RAB34对体外肿瘤发展的影响。
结果：在NSCLC组织和细胞中，PVT1和RAB34上调，miR-148下调。PVT1的过表达能够促进NSCLC细胞增殖和迁移，这可以通过miR-148恢复或RAB34敲低来逆转。此外，我们的数据首先确定RAB34的下调具有抑制作用,而RAB34的上调在体外和体内对肿瘤生长具有促进作用.
结论：这些结果表明，信号通路PVT1/miR-148/RAB34在NSCLC的进展中起关键作用，可作为NSCLC的可能诊断或治疗靶点。
