He Saifei, Ma Xing, Ye Ying, Zhang Miao, Zhuang Juhua, Song Yanan, Xia Wei
Central Laboratory, Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai, People's Republic of China.
Department of Nuclear Medicine, Seventh People's Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai, People's Republic of China.
Onco Targets Ther. 2019 May 21;12:4001-4011. doi: 10.2147/OTT.S195826. eCollection 2019.
To determine the mechanisms of HEATR1 on cell survival in non-small cell lung cancer (NSCLC). HEATR1 mRNA expression levels in 57 pairs of NSCLC tumor and adjacent normal lung tissues were analyzed using the TCGA database. The effect of HEATR1 inhibition on cell proliferation, apoptosis, and colony formation was measured in A549 and NCI-H460 cells lines. In addition, the effect of HEATR1 inhibition on tumor growth was measured using in vivo xenograft nude mouse models. Additionally, downstream signaling pathways affected by HEATR1 inhibition were analyzed using microarrays and bioinformatics analysis, and were validated using quantitative real-time polymerase chain reaction and Western blot analysis. HEATR1 levels were significantly higher in NSCLC tumor tissues compared to normal adjacent lung tissues (<0.001). In vitro, cell proliferation was significantly reduced in both A549 and NCI-H1299 cells transduced with shHEATR1 compared to shCtrl (<0.001). Colony formation was also significantly reduced after HEATR1 interference (<0.01). Additionally, the percentage of apoptosis was significantly increased in cells transduced with shHEATR1 (<0.001). In vivo, HEATR1 inhibition significantly reduced xenograft tumor growth in nude mice. HEATR1 inhibition drastically affected the p53-signaling pathway, significantly up-regulating PUMA and BAX both at the mRNA and protein levels (<0.001), while BCL2 levels were significantly down-regulated (<0.01). The cell proliferation and apoptosis were recovered in cell transduced with shHEATR1 and shp53 compared to shHEATR1 (<0.05). HEATR1 inhibition activated p53 by reducing ribosome biogenesis, which subsequently led to NSCLC cell apoptosis and reduced cell survival through the p53-PUMA-BAX/BCL2 axis. Our results provide a mechanism by which therapeutic modulation of HEATR1 could be a treatment strategy for NSCLC. In addition, HEATR1 could be used as a potential biomarker for the prognosis or therapeutic evaluation of NSCLC.
为确定HEATR1在非小细胞肺癌（NSCLC）细胞存活中的作用机制。利用TCGA数据库分析了57对NSCLC肿瘤组织和邻近正常肺组织中HEATR1 mRNA的表达水平。在A549和NCI-H460细胞系中检测了HEATR1抑制对细胞增殖、凋亡和集落形成的影响。此外，利用体内异种移植裸鼠模型检测了HEATR1抑制对肿瘤生长的影响。另外，使用微阵列和生物信息学分析来分析受HEATR1抑制影响的下游信号通路，并通过定量实时聚合酶链反应和蛋白质免疫印迹分析进行验证。与邻近正常肺组织相比，NSCLC肿瘤组织中的HEATR1水平显著更高（<0.001）。在体外，与转导shCtrl的细胞相比，转导shHEATR1的A549和NCI-H1299细胞的细胞增殖均显著降低（<0.001）。HEATR1干扰后集落形成也显著减少（<0.01）。此外，转导shHEATR1的细胞凋亡百分比显著增加（<0.001）。在体内，HEATR1抑制显著降低了裸鼠体内异种移植肿瘤的生长。HEATR1抑制极大地影响了p53信号通路，在mRNA和蛋白质水平上均显著上调PUMA和BAX（<0.001），而BCL2水平显著下调（<0.01）。与转导shHEATR1的细胞相比，转导shHEATR1和shp53的细胞的细胞增殖和凋亡得到恢复（<0.05）。HEATR1抑制通过减少核糖体生物合成来激活p53，随后通过p53-PUMA-BAX/BCL2轴导致NSCLC细胞凋亡并降低细胞存活率。我们的研究结果提供了一种机制，通过该机制对HEATR1进行治疗性调节可能成为NSCLC的一种治疗策略。此外，HEATR1可作为NSCLC预后或治疗评估的潜在生物标志物。
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