环状RNA UBAP2沉默对肺癌A549细胞体外增殖和侵袭的影响及机制

发布时间：2018-05-19 03:23

本文选题：circUBAP + 肺肿瘤　；参考：《中国肺癌杂志》2017年12期

【摘要】：背景与目的已有的研究证明环状RNA(circular RNAs,circ RNAs)在多种肿瘤发生发展中起重要作用,circUBAP2是一种促癌的circ RNA,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。
[Abstract]:Background & objective\ RNA(circular\ RNA(circular\ The aim of this study was to investigate the effect of circular RNA UBAP2circUBAP2 on proliferation and invasion of human lung cancer A549 cells in vitro and its mechanism. Methods CCK-8 assay was used to detect the inhibitory effect of circUBAP2 expression silencing on the growth of A549 cells. The effect of circUBAP2 expression silencing on the cell cycle and apoptosis of A549 cells was detected by flow cytometry. The effects of circUBAP2 expression silencing on the invasive ability of A549 cells were detected by Transwell assay. Western blot and Real-time PCR were used to detect the expression changes of JNK and ERK1/2 in A549 cells. Luciferase report technique was used to detect circUBAP2 direct target gene. Results the results of CCK-8 assay showed that the proliferation ability of A549 cell line was inhibited after the silencing of circUBAP2 expression, and the cell cycle stopped at G0 / G1 phase after the silencing of circUBAP2 expression, and the apoptosis rate increased significantly. The expression level of CDK6 cyclin D1c-IAP1C Bcl-2survivin FAKA Rac1MMP2 was significantly decreased, while the expression of p27 and Bax was significantly increased after silencing the expression of CDK6 cyclin D1nc-IAP1, the results of Western blot and Real-time PCR showed that the expression of CDK6 cyclin D1c-IAP1 and survivin FAK1 / MMP2 was significantly decreased, but the expression of p27 and Bax was significantly increased. Moreover, the activity of JNKN ERK / 1 / 2 was also significantly inhibited by the direct target of circUBAP2 because of miR-339-5pnmiR-96-3p and miR-135b-3p. Conclusion circUBAP2 plays an important role in the proliferation and invasion of lung cancer in vitro and inhibits its expression. It is expected to be a new target of molecular targeted therapy for lung cancer.
【作者单位】：包头肿瘤医院病理科;
【分类号】：R734.2

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