Nicotine-mediated invasion and migration of non-small cell lung carcinoma cells by modulating STMN3 and GSPT1 genes in an ID1-dependent manner.
Nicotine-mediated invasion and migration of non-small cell lung carcinoma cells by modulating STMN3 and GSPT1 genes in an ID1-dependent manner.
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DOI：
10.1186/1476-4598-13-173
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发表时间：
2014-07-16
期刊：
影响因子：
37.3
通讯作者：
Chellappan S
中科院分区：
文献类型：
--
作者：
Nair S;Bora-Singhal N;Perumal D;Chellappan S
DNA结合/分化抑制因子1（Inhibitor of DNA binding/Differentiation 1，ID 1）是一种缺乏基本DNA结合结构域的螺旋环螺旋转录因子。ID 1的过表达与多种人类癌症相关;我们早期的研究表明，已报道的ID 1由尼古丁或EGF刺激非小细胞肺癌（NSCLC）细胞诱导，其下调消除了细胞增殖，侵袭和迁移。在这里，我们试图确定介导这些效应的ID 1的下游靶点。对两种不同的NSCLC细胞系（A549和H1650）进行微阵列分析，所述细胞系用针对ID 1的siRNA或对照非靶向siRNA转染。用尼古丁刺激细胞，分析尼古丁刺激和ID 1消耗后差异表达的基因，以鉴定ID 1的潜在下游靶标。通过RT-PCR验证了所鉴定基因的潜在作用。进行额外的功能测定以评估这些基因在尼古丁诱导的增殖、侵袭和迁移中的作用。实验还阐明了不直接与DNA结合的ID 1在转录水平上影响这些基因表达的作用。微阵列分析显示，多个基因受到ID 1耗尽的影响;我们专注于其中两个：Stathmin样3（STMN 3），一种微管不稳定蛋白，和GSPT 1，一种参与翻译终止的蛋白质;这些蛋白质由尼古丁和EGF以ID 1依赖的方式诱导。ID 1在两种不同细胞系中的过表达在转录水平上诱导STMN 3和GSPT 1，而ID 1的缺失降低了它们的表达。STMN 3和GSPT 1被发现促进NSCLC细胞响应于nAChR活化的增殖、侵袭和迁移。试图评估ID 1，这是一种转录阻遏物，如何诱导这些基因表明，ID 1下调两个转录辅阻遏物，NRSF和ZBP 89，参与这些基因的抑制的表达。总的来说，我们的数据表明，尼古丁和EGF诱导基因，如STMN 3和GSPT 1，以促进非小细胞肺癌的增殖，侵袭和迁移，从而增强其致瘤性。因此，这些研究揭示了ID 1及其下游靶点在促进肺癌进展中的核心作用。
Inhibitor of DNA binding/Differentiation 1 (ID1) is a helix loop helix transcription factor that lacks the basic DNA binding domain. Over-expression of ID1 has been correlated with a variety of human cancers; our earlier studies had shown that reported ID1 is induced by nicotine or EGF stimulation of non-small cell lung cancer (NSCLC) cells and its down regulation abrogates cell proliferation, invasion and migration. Here we made attempts to identify downstream targets of ID1 that mediate these effects. A microarray analysis was done on two different NSCLC cell lines (A549 and H1650) that were transfected with a siRNA to ID1 or a control, non-targeting siRNA. Cells were stimulated with nicotine and genes that were differentially expressed upon nicotine stimulation and ID1 depletion were analyzed to identify potential downstream targets of ID1. The prospective role of the identified genes was validated by RT-PCR. Additional functional assays were conducted to assess the role of these genes in nicotine induced proliferation, invasion and migration. Experiments were also conducted to elucidate the role of ID1, which does not bind to DNA directly, affects the expression of these genes at transcriptional level. A microarray analysis showed multiple genes are affected by the depletion of ID1; we focused on two of them: Stathmin-like3 (STMN3), a microtubule destabilizing protein, and GSPT1, a protein involved in translation termination; these proteins were induced by both nicotine and EGF in an ID1 dependent fashion. Overexpression of ID1 in two different cell lines induced STMN3 and GSPT1 at the transcriptional level, while depletion of ID1 reduced their expression. STMN3 and GSPT1 were found to facilitate the proliferation, invasion and migration of NSCLC cells in response to nAChR activation. Attempts made to assess how ID1, which is a transcriptional repressor, induces these genes showed that ID1 down regulates the expression of two transcriptional co-repressors, NRSF and ZBP89, involved in the repression of these genes. Collectively, our data suggests that nicotine and EGF induce genes such as STMN3 and GSPT1 to promote the proliferation, invasion and migration of NSCLC, thus enhancing their tumorigenic properties. These studies thus reveal a central role for ID1 and its downstream targets in facilitating lung cancer progression.
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