Armas-López Leonel, Piña-Sánchez Patricia, Arrieta Oscar, de Alba Enrique Guzman, Ortiz-Quintero Blanca, Santillán-Doherty Patricio, Christiani David C, Zúñiga Joaquín, Ávila-Moreno Federico
National University Autonomous of México (UNAM), Facultad de Estudios Superiores (FES) Iztacala, Biomedicine Research Unit (UBIMED), Lung Diseases And Cancer Epigenomics Laboratory, Mexico State, Mexico.
Instituto Mexicano del Seguro Social (IMSS), Centro Medico Nacional (CMN) Siglo XXI, Unidad de Investigación Médica en Enfermedades Oncológicas (UIMEO), Molecular Oncology Laboratory, Mexico City, Mexico.
Oncotarget. 2017 May 9;8(40):67056-67081. doi: 10.18632/oncotarget.17715. eCollection 2017 Sep 15.
Several homeobox-related gene (HOX) transcription factors such as mesenchyme HOX-2 (MEOX2) have previously been associated with cancer drug resistance, malignant progression and/or clinical prognostic responses in lung cancer patients; however, the mechanisms involved in these responses have yet to be elucidated. Here, an epigenomic strategy was implemented to identify novel MEOX2 gene promoter transcription targets and propose a new molecular mechanism underlying lung cancer drug resistance and poor clinical prognosis. Chromatin immunoprecipitation (ChIP) assays derived from non-small cell lung carcinomas (NSCLC) hybridized on gene promoter tiling arrays and bioinformatics analyses were performed, and quantitative, functional and clinical validation were also carried out. We statistically identified a common profile consisting of 78 gene promoter targets, including Hedgehog-GLI1 gene promoter sequences (FDR≤0.1 and FDR≤0.2). The GLI-1 gene promoter region from -2,192 to -109 was occupied by MEOX2, accompanied by transcriptionally active RNA Pol II and was epigenetically linked to the active histones H3K27Ac and H3K4me3; these associations were quantitatively validated. Moreover, siRNA genetic silencing assays identified a MEOX2-GLI1 axis involved in cellular cytotoxic resistance to cisplatinum in a dose-dependent manner, as well as cellular migration and proliferation. Finally, Kaplan-Maier survival analyses identified significant MEOX2-dependent GLI-1 protein expression associated with clinical progression and poorer overall survival using an independent cohort of NSCLC patients undergoing platinum-based oncological therapy with both epidermal growth factor receptor (EGFR)-non-mutated and EGFR-mutated status. In conclusion, this is the first study to investigate epigenome-wide MEOX2-transcription factor occupation identifying a novel overexpressed MEOX2-GLI1 axis and its clinical association with platinum-based cancer drug resistance and EGFR-tyrosine kinase inhibitor (TKI)-based therapy responses in NSCLC patients.
几种与同源框相关的基因(HOX)转录因子,如间充质HOX-2(MEOX2),此前已被发现与肺癌患者的癌症耐药性、恶性进展和/或临床预后反应有关;然而,这些反应所涉及的机制尚未阐明。在此,我们实施了一种表观基因组策略,以识别新的MEOX2基因启动子转录靶点,并提出一种新的分子机制,解释肺癌耐药性和不良临床预后的原因。我们进行了源自非小细胞肺癌(NSCLC)并与基因启动子平铺阵列杂交的染色质免疫沉淀(ChIP)分析以及生物信息学分析,同时也进行了定量、功能和临床验证。我们通过统计学方法确定了一个由78个基因启动子靶点组成的共同图谱,其中包括刺猬索尼信号通路相关蛋白1(Hedgehog-GLI1)基因启动子序列(错误发现率≤0.1和错误发现率≤0.2)。从-2192到-109的GLI-1基因启动子区域被MEOX2占据,同时伴有转录活性的RNA聚合酶II,并且在表观遗传上与活性组蛋白H3K27Ac和H3K4me3相关联;这些关联通过定量分析得到了验证。此外,小干扰RNA(siRNA)基因沉默分析确定了MEOX2-GLI1轴以剂量依赖的方式参与细胞对顺铂的细胞毒性抗性,以及细胞迁移和增殖。最后,通过Kaplan-Meier生存分析,在一组接受铂类肿瘤治疗的非小细胞肺癌患者独立队列中,无论其表皮生长因子受体(EGFR)是否突变,均发现显著的MEOX2依赖性GLI-1蛋白表达与临床进展和较差的总生存率相关。总之,这是第一项研究全表观基因组范围内MEOX2转录因子占据情况的研究,确定了一个新的过表达的MEOX2-GLI1轴及其与非小细胞肺癌患者铂类癌症耐药性和基于表皮生长因子受体酪氨酸激酶抑制剂(TKI)治疗反应的临床关联。