Upregulated lncRNA ADAMTS9-AS2 suppresses progression of lung cancer through inhibition of miR-223-3p and promotion of TGFBR3
Upregulated lncRNA ADAMTS9-AS2 suppresses progression of lung cancer through inhibition of miR-223-3p and promotion of TGFBR3
复制标题
上调的 lncRNA ADAMTS9-AS2 通过抑制 miR-223-3p 和促进 TGFBR3 抑制肺癌进展
DOI：
10.1002/iub.1752
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发表时间：
2018-06-01
期刊：
影响因子：
4.6
通讯作者：
Chen, Ting
中科院分区：
文献类型：
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作者：
Liu, Chao;Yang, Zuozhang;Chen, Ting
在本研究中，我们旨在通过调节miR-223-3p和TGFBR3的表达来研究lncRNA ADAMTS9-AS2在肺癌进展中的作用。用逆转录聚合酶链式反应(qRT-PCR)检测ADAMTS9-AS2在肺癌组织和细胞系中的表达。分别使用TargetScan和miRcode预测靶向关系。用荧光素酶报告系统验证了ADAMTS9-AS2、TGFBR3和miR-223-3p之间的关系。Western印迹法检测TGFBR3蛋白水平的变化。CCK-8比色法测定细胞增殖。流式细胞仪检测细胞周期和细胞凋亡率，Transwell法检测细胞迁移和侵袭能力。建立肿瘤移植模型，研究ADAMTS9-AS2对体内肿瘤生长的影响。QRT-PCR结果显示，lncADAMTS9-AS2在肺癌组织中低表达。ADAMTS9-AS2在肺癌细胞中的高表达显著降低了细胞的增殖能力，抑制了细胞的迁移，并提高了细胞的凋亡率。体内实验发现ADAMTS9-AS2对肺癌的生长有抑制作用。生物信息学预测miR-223-3p直接与ADAMTS9-AS2和TGFBR3结合，后来被荧光素酶报告系统证实。ADAMTS9-AS2基因表达上调，而miR-223-3p基因表达显著降低。此外，我们的结果显示miR-223-3p诱导细胞凋亡，而TGFBR3组表现出完全相反的作用。证实ADAMTS9-AS2和TGFBR3是miR-223-3p的直接基因。MIR-223-3p通过靶向TGFBR3促进肺癌细胞的增殖、迁移和侵袭。因此，ADAMTS9-AS2、miR-223-3p和TGFBR3可能成为肺癌治疗的潜在靶点。(C)2018年IUBMB Life，70(6)：536-546,2018
In this study, we aimed at investigating effects of lncRNA ADAMTS9-AS2 on lung cancer progression through regulating miR-223-3p and TGFBR3 expressions. Expressions of ADAMTS9-AS2 in lung cancer tissues and cell lines were determined by reverse transcriptase polymerase chain reaction (qRT-PCR). TargetScan and miRcode were used to predict the targeting relationships, respectively. The luciferase reporter system was used to verify that the relationship among ADAMTS9-AS2, TGFBR3 and miR-223-3p. Western blot assay tested the protein level changes in TGFBR3. Cell proliferation was determined by CCK-8 assay. Cell cycle and cell apoptosis were detected by flow cytometry assay, and migration and invasion were determined by transwell assay. Tumor xenograft model was developed to study the influence of ADAMTS9-AS2 on tumor growth in vivo. qRT-PCR results demonstrated that lncADAMTS9-AS2 was lowly expressed in lung cancer tissues. High expression of ADAMTS9-AS2 in lung cancer cells significantly reduced proliferation ability and inhibited migration, as well as elevating their apoptosis rate. In vivo assay found that ADAMTS9-AS2 suppressed the lung tumor growth. Bioinformatics predicted that miR-223-3p bound directly to the ADAMTS9-AS2 and TGFBR3, which was later confirmed by luciferase reporter system. ADAMTS9-AS2 transfection increased TGFBR3 mRNA and protein expressions in lung cancer cells, but miR-223-3p transfection significantly decreased them. Besides, our results showed that miR-223-3p induced cellular apoptosis while TGFBR3 group showed the complete opposite effect. It was proved that ADAMTS9-AS2 and TGFBR3 were the direct genes of miR-223-3p. MiR-223-3p promotes proliferation, migration and invasion of lung cancer cells by targeting TGFBR3. Therefore, ADAMTS9-AS2, miR-223-3p and TGFBR3 may provide potential targets for the treatment of lung cancer patients. (c) 2018 IUBMB Life, 70(6):536-546, 2018