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環(huán)狀RNA UBAP2沉默對肺癌A549細(xì)胞體外增殖和侵襲的影響及機(jī)制

發(fā)布時間:2018-05-19 03:23

  本文選題:circUBAP + 肺腫瘤; 參考:《中國肺癌雜志》2017年12期


【摘要】:背景與目的已有的研究證明環(huán)狀RNA(circular RNAs,circ RNAs)在多種腫瘤發(fā)生發(fā)展中起重要作用,circUBAP2是一種促癌的circ RNA,但是其在肺癌中的作用和機(jī)制尚不明確。本研究旨在探討circular RNA UBAP2(circUBAP2)對人肺癌A549細(xì)胞體外增殖和侵襲能力的影響并初步探討其機(jī)制。方法應(yīng)用CCK-8法檢測circUBAP2表達(dá)沉默對A549細(xì)胞生長的抑制作用,采用流式細(xì)胞儀技術(shù)檢測circUBAP2表達(dá)沉默對A549細(xì)胞周期和細(xì)胞失巢凋亡的影響,Transwell實驗檢測circUBAP2表達(dá)沉默對A549細(xì)胞侵襲能力的影響,Western blot和Real-time PCR檢測CDK6、cyclin D1、p27、c-IAP1、Bcl-2、Survivin、Bax、FAK、Rac1、MMP2的表達(dá)變化以及JNK和ERK1/2的活性,熒光素酶報告技術(shù)檢測circUBAP2直接靶基因。結(jié)果 CCK-8實驗結(jié)果顯示circUBAP2表達(dá)沉默后,A549細(xì)胞株增殖能力受抑制;流式細(xì)胞結(jié)果顯示,circUBAP2表達(dá)沉默后,細(xì)胞周期停滯在G0期/G1期,且細(xì)胞凋亡率顯著增加;Transwell實驗結(jié)果顯示,細(xì)胞體外侵襲能力明顯受到抑制。Western blot和Real-time PCR結(jié)果顯示,circUBAP2表達(dá)沉默后,CDK6、cyclin D1、c-IAP1、Bcl-2、Survivin、FAK、Rac1、MMP2等基因的表達(dá)水平顯著下降,而p27和Bax的表達(dá)顯著上升,而且JNK、ERK1/2的活性也受到明顯的抑制,circUBAP2直接靶基因為miR-339-5p、miR-96-3p和miR-135b-3p。結(jié)論 circUBAP2在肺癌的體外增殖和侵襲中發(fā)揮重要的作用,抑制其表達(dá),有望成為肺癌分子靶向治療的新靶點。
[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.
【作者單位】: 包頭腫瘤醫(yī)院病理科;
【分類號】:R734.2

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