Induction of long intergenic non-coding RNA HOTAIR in lung cancer cells by type I collagen
- Yan Zhuang†1,
- Xiang Wang†2,
- Hong T Nguyen1,
- Ying Zhuo1,
- Xinpeng Cui2,
- Claire Fewell1,
- Erik K Flemington1 and
- Bin Shan1, 3Email author
© Zhuang et al.; licensee BioMed Central Ltd. 2013
Received: 18 March 2013
Accepted: 7 May 2013
Published: 13 May 2013
The tumor microenvironment is a crucial determinant in tumor progression. Interstitial extracellular matrix (ECM), such as type I collagen (Col-1), is aberrantly enriched in the tumor microenvironment and promotes tumor progression. Long intergenic non-coding RNAs (lincRNA) are a new family of regulatory RNAs that modulate fundamental cellular processes via diverse mechanisms.
We investigated whether the expression of lincRNAs was regulated by the tumor promoting Col-1. In a three-dimensional organotypic culture model using the reconstituted basement membrane ECM Matrigel (rBM 3-D), supplementation of Col-1 disrupted acini, a differentiation feature of well-differentiated lung adenocarcinoma cells, and concurrently induced the expression of a tumor-promoting lincRNA, HOX transcript antisense RNA (HOTAIR). Induction of HOTAIR by Col-1 was diminished by a neutralizing antibody against the Col-1 receptor α2β1 integrin. Col-1 activates the expression of a reporter gene controlled by the human HOTAIR promoter. Moreover the expression of HOTAIR and Col-1 was concurrently up-regulated in human non-small cell lung cancer.
Our findings indicate that tumor-promoting Col-1 up-regulates the expression of HOTAIR in NSCLC cells. These initial results warrant further investigation of HOTAIR and other lincRNA genes in lung tumorigenesis.
KeywordsHOTAIR lincRNA Type I collagen Three-dimensional ogranotypic culture
The tumor microenvironment is aberrantly enriched with interstitial extracellular matrix (ECM), such as type I collagen (Col-1) . The tumor-promoting activity of Col-1 has been successfully investigated using three-dimensional organotypic culture based on reconstituted basement membrane matrix Matrigel (rBM 3-D) . In rBM 3-D culture, normal lung and mammary epithelial cells form acini, a differentiation feature that manifests spheres of polarized epithelial cells enclosing a central lumen [3, 4]. Col-1 disrupts acinar morphogenesis of lung and mammary epithelial cells in rBM 3-D culture and promotes tumor progression in vivo[5, 6]. Moreover, Col-1 up-regulates the expression of several oncogenic miRNAs, such as miR-21 in rBM 3-D culture of lung and mammary epithelial cells [6, 7].
Long intergenic non-coding RNAs (lincRNAs) are a new family of regulatory RNAs that modulate tumorigenesis via diverse mechanisms . HOX transcript antisense RNA (HOTAIR) is a HOXC cluster-derived lincRNA that binds to the transcriptional co-repressor polycomb repressive complex 2 (PRC2) and recruits PRC2 to silence its target genes . HOTAIR is proposed as an oncogene because its expression is elevated in several types of cancers and mediates invasion and metastasis of breast cancer cells [10–12]. Regulation of the expression of HOTAIR in cancer remains unclear and is investigated in the current study.
In the current study, we demonstrate that the tumor-promoting lincRNA HOTAIR is induced by Col-1 and its expression inversely correlates with acinar morphogenesis, a differentiation feature of lung epithelial cells in rBM 3-D culture (Figure 1) . These in vitro findings suggest that the elevated HOTAIR expression in tumor tissues results from cancer cells’ response to Col-1 that is aberrantly enriched in the tumor microenvironment (Figure 3) [2, 5, 19]. Col-1 appears to transcriptionally activate the HOTAIR gene because Col-1 activates a reporter gene controlled by the human HOTAIR promoter (Figure 2). In silico analysis of the human HOTAIR promoter using the geWorkbench Promoter Scanning Module reveals four potential Myc binding sites (https://wiki.nci.nih.gov/display/geWorkbench/geWorkbench) (Additional file 1: Table S3). It is conceivable that Myc mediates activation of the HOTAIR gene by Col-1 because the miR-17-92 cluster, another transcriptional target of Myc in cancer cells, is concurrently up-regulated by Col-1 in rBM 3-D culture in our recent report [6, 24]. The clinical relevance of our findings is supported by a concurrent up-regulation of HOTAIR and Col-1 expression in NSCLC (Figure 3). A lack of quantitative correlation between HOTAIR and Col-1 expression implicates additional signaling in regulation of the expression of HOTAIR in NSCLC. Because the specimens were collected recently, it is unclear whether the elevated expression of HOTAIR holds prognostic values in NSCLC as in several other types of tumors [10–12]. The patients are enrolled in a follow-up study to monitor progression and survival.
To the best of our knowledge, the current study is the very first attempt to examine a link between the tumor microenvironment and lincRNAs using rBM 3-D organotypic culture. Because HOTAIR can epigenetically reprogram global chromatin staging and gene expression via its interaction with PRC2, our initial findings warrant further investigation of HOTAIR-mediated epigenetic regulation of the crosstalk between cancer cells and their microenvironment.
Type I collagen
Long intergenic non-coding RNA
HOX transcript antisense RNA
- rBM 3-D:
Reconstituted basement membrane matrix Matrigel based three-dimensional culture
Polycomb repressive complex 2
Non-small cell lung cancer.
This work is supported by Tulane Research Enhancement Fund awarded to BS.
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