Chin, SP and Marthick, JR and West, AC and Short, AK and Chuckowree, J and Polanowski, AM and Thomson, RJ and Holloway, AF and Dickinson, JL, Regulation of the ITGA2 gene by epigenetic mechanisms in prostate cancer, The Prostate, 75, (7) pp. 723-734. ISSN 0270-4137 (2015) [Refereed Article]
Copyright 2015 Wiley Periodicals, Inc.
Background: Integrin alpha2 beta1 (α2β1) plays an integral role in tumour cell invasion, metastasis and angiogenesis, and altered expression of the receptor has been linked to tumour prognosis in several solid tumours. However, the relationship is complex, with both increased and decreased expression associated with different stages of tumour metastases in several tumour types. The ITGA2 gene, which codes for the α2 subunit, was examined to investigate whether a large CpG island associated with its promoter region is involved in the differential expression of ITGA2 observed in prostate cancer.
Methods: Bisulphite sequencing of the ITGA2 promoter was used to assess methylation in formalin-fixed paraffin-embedded (FFPE) prostate tumour specimens and prostate cancer cell lines, PC3, 22Rv1 and LNCaP. Changes in ITGA2 mRNA expression were measured using quantitative PCR. ITGA2 functionality was interrogated using cell migration scratch assays and siRNA knockdown experiments.
Results: Bisulphite sequencing revealed strikingly decreased methylation at key CpG sites within the promoter of tumour samples, when compared with normal prostate tissue. Altered methylation of this CpG island is also associated with differences in expression in the non-invasive LNCaP, and the highly metastatic PC3 and 22Rv1 prostate cancer cell lines. Further bisulphite sequencing confirmed that selected CpGs were highly methylated in LNCaP cells, whilst only low levels of methylation were observed in PC3 and 22Rv1 cells, correlating with ITGA2 transcript levels. Examination of the increased expression of ITGA2 was shown to influence migratory potential via scratch assay in PC3, 22Rv1 and LNCaP cells, and was confirmed by siRNA knockdown experiments.
Conclusions: Taken together, our data supports the assertion that epigenetic modification of the ITGA2 promoter is a mechanism by which ITGA2 expression is regulated.
|Item Type:||Refereed Article|
|Keywords:||integrins, DNA methylation, prostate, epigenetics|
|Research Division:||Biological Sciences|
|Research Field:||Epigenetics (incl. genome methylation and epigenomics)|
|Objective Group:||Clinical health|
|Objective Field:||Clinical health not elsewhere classified|
|UTAS Author:||Chin, SP (Miss Suyin Chin)|
|UTAS Author:||Marthick, JR (Mr James Marthick)|
|UTAS Author:||West, AC (Ms Alison West)|
|UTAS Author:||Short, AK (Ms Annabel Short)|
|UTAS Author:||Chuckowree, J (Dr Jyoti Chuckowree)|
|UTAS Author:||Polanowski, AM (Ms Andrea Polanowski)|
|UTAS Author:||Thomson, RJ (Dr Russell Thomson)|
|UTAS Author:||Holloway, AF (Professor Adele Holloway)|
|UTAS Author:||Dickinson, JL (Professor Joanne Dickinson)|
|Web of Science® Times Cited:||16|
|Deposited By:||Menzies Institute for Medical Research|
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