Clinical Laboratory, Shanghai Pudong New Area Gongli Hospital, Shanghai, China.
Department of Emergency Medicine, Shanghai Pudong New Area Gongli Hospital, The Second Military Medical University, Shanghai, China.
J Cell Biochem. 2019 Nov;120(11):19019-19030. doi: 10.1002/jcb.29225. Epub 2019 Jun 26.
Circular RNAs (circRNAs) have an important function in human diseases, especially in cancer. circRNA hsa_circ_0014130 (circPIP5K1A), a particularly abundant circRNA, participates in the tumorigenesis of non-small cell lung cancer (NSCLC), although the underlying regulatory mechanism remains unclear. Here, we investigated the circPIP5K1A role in NSCLC. Expression of circPIP5K1A in NSCLC cell lines was explored with quantitative real-time PCR. The effect of circPIP5K1A on NSCLC was evaluated with circPIP5K1A silencing, miR-600 mimic transfection, and hypoxia-inducible factor (HIF)-1α overexpression, followed by assessment of cell proliferation, metastasis, and tumorigenesis in nude mice. The subcellular localization of circPIP5K1A was evaluated via fluorescence in situ hybridization (FISH), and correlation between circPIP5K1A, miR-600, and HIF-1α was assessed by luciferase assay. The data demonstrated that circPIP5K1A expression was increased in NSCLC cells. FISH showed that circPIP5K1A localized to the cytoplasm. The circPIP5K1A knockdown suppressed NSCLC cell metastasis and proliferation by promoting expression of miR-600. Overexpression of miR-600 inhibited HIF-1α-mediated metastasis and proliferation of NSCLC cell by downregulating the endothelial mesenchymal transition-related proteins, Snail and vimentin, and upregulating E-cadherin. In vivo experiments illustrated that circPIP5K1A silence suppressed tumor growth and pulmonary metastasis. The circPIP5K1A may function as an miR-600 sponge to facilitate NSCLC proliferation and metastasis by promoting HIF-1α. A bifluorescein reporter experiment confirmed that miR-600 was the circPIP5K1A target, and miR-600 interacted with the 3' untranslated region of HIF-1α. These results show that circPIP5K1A acted as a tumor promoter through a novel circPIP5K1A/miR-600/HIF-1α axis, which provides candidate markers and therapeutic targets for NSCLC.
环状 RNA（circRNAs）在人类疾病中具有重要功能，特别是在癌症中。环状 RNA hsa_circ_0014130（circPIP5K1A）是一种特别丰富的环状 RNA，参与非小细胞肺癌（NSCLC）的肿瘤发生，尽管其潜在的调控机制尚不清楚。在这里，我们研究了 circPIP5K1A 在 NSCLC 中的作用。通过定量实时 PCR 探索 NSCLC 细胞系中 circPIP5K1A 的表达。通过 circPIP5K1A 沉默、miR-600 模拟物转染和缺氧诱导因子（HIF）-1α过表达评估 circPIP5K1A 对 NSCLC 的影响，然后在裸鼠中评估细胞增殖、转移和肿瘤发生。通过荧光原位杂交（FISH）评估 circPIP5K1A 的亚细胞定位，并通过荧光素酶测定评估 circPIP5K1A、miR-600 和 HIF-1α 之间的相关性。数据表明，circPIP5K1A 在 NSCLC 细胞中表达增加。FISH 显示 circPIP5K1A 定位于细胞质。circPIP5K1A 敲低通过促进 miR-600 的表达抑制 NSCLC 细胞转移和增殖。miR-600 的过表达通过下调内皮间质转化相关蛋白 Snail 和波形蛋白并上调 E-钙黏蛋白抑制 HIF-1α 介导的 NSCLC 细胞转移和增殖。体内实验表明，circPIP5K1A 沉默抑制肿瘤生长和肺转移。circPIP5K1A 可能作为 miR-600 的海绵通过促进 HIF-1α 来促进 NSCLC 的增殖和转移。双荧光素酶报告实验证实 miR-600 是 circPIP5K1A 的靶标，miR-600 与 HIF-1α 的 3'UTR 相互作用。这些结果表明，circPIP5K1A 通过新型 circPIP5K1A/miR-600/HIF-1α 轴作为肿瘤促进因子发挥作用，为 NSCLC 提供了候选标志物和治疗靶点。
