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Long Non-Coding RNA TRG-AS1 Promoted Proliferation and Invasion of Lung Cancer Cells Through the miR-224-5p/SMAD4 Axis
OncoTargets and Therapy ( IF 2.7 ) Pub Date : 2021-08-10 , DOI: 10.2147/ott.s297336 Mengyan Zhang 1, 2 , Weiguo Zhu 3 , Mansour Haeryfar 4 , Sumei Jiang 5 , Xiang Jiang 6 , Wei Chen 7 , Jiancheng Li 2
OncoTargets and Therapy ( IF 2.7 ) Pub Date : 2021-08-10 , DOI: 10.2147/ott.s297336 Mengyan Zhang 1, 2 , Weiguo Zhu 3 , Mansour Haeryfar 4 , Sumei Jiang 5 , Xiang Jiang 6 , Wei Chen 7 , Jiancheng Li 2
Affiliation
Introduction: The aim of this study was to investigate the role and mechanism of long non-coding RNA (lncRNA) TRG-AS1 in mediating the proliferation, invasion and migration of lung cancer cells as well lung tumor growth.
Methods: Firstly, the expression levels of TRG-AS1, miR-224-5p in lung cancer tissues or cells were quantified by quantitative real-time PCR. Western blot analysis was conducted to measure the expression levels of protein SMAD4. CCK-8 assay, wound healing assay and transwell assay were conducted to evaluate cell proliferation, migration and invasion, respectively. The interaction between TRG-AS1 and miR-224-5p was predicted by bioinformatics analysis. Dual-luciferase assay and RNA pull-down assay were performed to further confirm their interaction. In addition, the interaction between miR-224-5p and SMAD4 was detected by RIP assay.
Results: The results showed that TRG-AS1 was highly upregulated and miR-224-5p was downregulated in lung cancer. A negative correlation was found between TRG-AS1 and miR-224-5p. Furthermore, upregulation of TRG-AS1 promoted cell proliferation and invasion, while overexpression of miR-224-5p attenuated the effects of TRG-AS1. The downstream protein SMAD4 played an important role. In vivo study showed that knockdown of TRG-AS1 effectively retarded tumor growth.
Discussion: Our data suggested that the TRG-AS1/miR-224-5p/SMAD4 axis may be a potential therapeutic target in lung cancer.
Keywords: lung cancer, TRG-AS1, miR-224-5p/SMAD4 axis, therapeutic target
中文翻译：
长非编码RNA TRG-AS1通过miR-224-5p/SMAD4轴促进肺癌细胞增殖和侵袭
简介：本研究旨在探讨长链非编码RNA（lncRNA）TRG-AS1在介导肺癌细胞增殖、侵袭和迁移以及肺肿瘤生长中的作用和机制。
方法：首先采用实时荧光定量PCR法检测肺癌组织或细胞中TRG-AS1、miR-224-5p的表达水平。进行蛋白质印迹分析以测量蛋白质SMAD4的表达水平。 CCK-8实验、伤口愈合实验和Transwell实验分别评估细胞增殖、迁移和侵袭。通过生物信息学分析预测TRG-AS1和miR-224-5p之间的相互作用。进行双荧光素酶测定和RNA Pull-down测定以进一步证实它们的相互作用。此外，通过RIP测定检测miR-224-5p和SMAD4之间的相互作用。
结果：结果显示，肺癌中TRG-AS1高度上调，miR-224-5p下调。 TRG-AS1 和 miR-224-5p 之间存在负相关。此外，TRG-AS1的上调促进细胞增殖和侵袭，而miR-224-5p的过度表达减弱TRG-AS1的作用。下游蛋白SMAD4发挥了重要作用。体内研究表明，TRG-AS1 的敲低可有效延缓肿瘤生长。
讨论：我们的数据表明 TRG-AS1/miR-224-5p/SMAD4 轴可能是肺癌的潜在治疗靶点。
关键词:肺癌, TRG-AS1, miR-224-5p/SMAD4轴, 治疗靶点
更新日期：2021-08-10
Methods: Firstly, the expression levels of TRG-AS1, miR-224-5p in lung cancer tissues or cells were quantified by quantitative real-time PCR. Western blot analysis was conducted to measure the expression levels of protein SMAD4. CCK-8 assay, wound healing assay and transwell assay were conducted to evaluate cell proliferation, migration and invasion, respectively. The interaction between TRG-AS1 and miR-224-5p was predicted by bioinformatics analysis. Dual-luciferase assay and RNA pull-down assay were performed to further confirm their interaction. In addition, the interaction between miR-224-5p and SMAD4 was detected by RIP assay.
Results: The results showed that TRG-AS1 was highly upregulated and miR-224-5p was downregulated in lung cancer. A negative correlation was found between TRG-AS1 and miR-224-5p. Furthermore, upregulation of TRG-AS1 promoted cell proliferation and invasion, while overexpression of miR-224-5p attenuated the effects of TRG-AS1. The downstream protein SMAD4 played an important role. In vivo study showed that knockdown of TRG-AS1 effectively retarded tumor growth.
Discussion: Our data suggested that the TRG-AS1/miR-224-5p/SMAD4 axis may be a potential therapeutic target in lung cancer.
Keywords: lung cancer, TRG-AS1, miR-224-5p/SMAD4 axis, therapeutic target
中文翻译：
长非编码RNA TRG-AS1通过miR-224-5p/SMAD4轴促进肺癌细胞增殖和侵袭
简介：本研究旨在探讨长链非编码RNA（lncRNA）TRG-AS1在介导肺癌细胞增殖、侵袭和迁移以及肺肿瘤生长中的作用和机制。
方法：首先采用实时荧光定量PCR法检测肺癌组织或细胞中TRG-AS1、miR-224-5p的表达水平。进行蛋白质印迹分析以测量蛋白质SMAD4的表达水平。 CCK-8实验、伤口愈合实验和Transwell实验分别评估细胞增殖、迁移和侵袭。通过生物信息学分析预测TRG-AS1和miR-224-5p之间的相互作用。进行双荧光素酶测定和RNA Pull-down测定以进一步证实它们的相互作用。此外，通过RIP测定检测miR-224-5p和SMAD4之间的相互作用。
结果：结果显示，肺癌中TRG-AS1高度上调，miR-224-5p下调。 TRG-AS1 和 miR-224-5p 之间存在负相关。此外，TRG-AS1的上调促进细胞增殖和侵袭，而miR-224-5p的过度表达减弱TRG-AS1的作用。下游蛋白SMAD4发挥了重要作用。体内研究表明，TRG-AS1 的敲低可有效延缓肿瘤生长。
讨论：我们的数据表明 TRG-AS1/miR-224-5p/SMAD4 轴可能是肺癌的潜在治疗靶点。
关键词:肺癌, TRG-AS1, miR-224-5p/SMAD4轴, 治疗靶点
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