Department of Nuclear Medicine, Tumor Hospital of Yunnan Province, The Third Affiliated Hospital of Kunming Medical College, Kunming, Yunnan, People's Republic of China.
Departments of Orthopaedics, Tumor Hospital of Yunnan Province, The Third Affiliated Hospital of Kunming Medical College, Kunming, Yunnan, People's Republic of China.
IUBMB Life. 2018 Jun;70(6):536-546. doi: 10.1002/iub.1752. Epub 2018 Apr 29.
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. © 2018 IUBMB Life, 70(6):536-546, 2018.
在这项研究中，我们旨在通过调节 lncRNA ADAMTS9-AS2 和 miR-223-3p 的表达来研究 lncRNA ADAMTS9-AS2 对肺癌进展的影响。通过逆转录聚合酶链反应（qRT-PCR）测定肺癌组织和细胞系中 ADAMTS9-AS2 的表达。分别使用 TargetScan 和 miRcode 预测靶向关系。使用荧光素酶报告系统验证 ADAMTS9-AS2、TGFBR3 和 miR-223-3p 之间的关系。Western blot 检测 TGFBR3 蛋白水平的变化。通过 CCK-8 测定法测定细胞增殖。通过流式细胞术检测细胞周期和细胞凋亡，通过 Transwell 测定法检测迁移和侵袭。建立肿瘤异种移植模型研究 ADAMTS9-AS2 对体内肿瘤生长的影响。qRT-PCR 结果表明，lncADAMTS9-AS2 在肺癌组织中低表达。肺癌细胞中 ADAMTS9-AS2 的高表达显著降低了增殖能力，并抑制了迁移，同时提高了细胞凋亡率。体内实验发现 ADAMTS9-AS2 抑制了肺肿瘤的生长。生物信息学预测 miR-223-3p 直接与 ADAMTS9-AS2 和 TGFBR3 结合，随后通过荧光素酶报告系统得到证实。ADAMTS9-AS2 转染增加了肺癌细胞中 TGFBR3 mRNA 和蛋白的表达，但 miR-223-3p 转染显著降低了它们的表达。此外，我们的结果表明，miR-223-3p 诱导细胞凋亡，而 TGFBR3 组则表现出完全相反的效果。证明 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 可能为肺癌患者的治疗提供潜在靶点。
