Abstract
背景与目的已有的研究证明环状RNA(circular RNAs, circRNAs)在多种肿瘤发生发展中起重要作用,circUBAP2是一种促癌的circRNA,但是其在肺癌中的作用和机制尚不明确。本研究旨在探讨circular RNA UBAP2(circUBAP2)对人肺癌A549细胞体外增殖和侵袭能力的影响并初步探讨其机制。方法应用CCK-8法检测circUBAP2表达沉默对A549细胞生长的抑制作用,采用流式细胞仪技术检测circUBAP2表达沉默对A549细胞周期和细胞失巢凋亡的影响,Transwell实验检测circUBAP2表达沉默对A549细胞侵袭能力的影响,Western blot和Real-time PCR检测CDK6、cyclin D1、p27、c-IAP1、Bcl-2、Survivin、Bax、FAK、Rac1、MMP2的表达变化以及JNK和ERK1/2的活性,荧光素酶报告技术检测circUBAP2直接靶基因。结果CCK-8实验结果显示circUBAP2表达沉默后,A549细胞株增殖能力受抑制;流式细胞结果显示,circUBAP2表达沉默后,细胞周期停滞在G0期/G1期,且细胞凋亡率显著增加;Transwell实验结果显示,细胞体外侵袭能力明显受到抑制。Western blot和Real-time PCR结果显示,circUBAP2表达沉默后,CDK6、cyclin D1、c-IAP1、Bcl-2、Survivin、FAK、Rac1、MMP2等基因的表达水平显著下降,而p27和Bax的表达显著上升,而且JNK、ERK1/2的活性也受到明显的抑制,circUBAP2直接靶基因为miR-339-5p、miR-96-3p和miR-135b-3p。结论circUBAP2在肺癌的体外增殖和侵袭中发挥重要的作用,抑制其表达,有望成为肺癌分子靶向治疗的新靶点。
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