Aberrant hypermethylation of ID4 gene promoter region increases risk of lymph node metastasis in T1 breast cancer

Abstract

ID4 gene is a member of the inhibitor of DNA-binding (ID) family, which inhibits DNA binding of basic helixloop-helix transcription factors. Certain human primary breast cancers reportedly have low or no expression of ID4 protein, but its role in carcinogenesis and cancer progression is unknown. To determine its possible role, we examined epigenetic inactivation of ID4 gene by promoter hypermethylation in human breast cell lines and T1 breast cancer tissues. Methylation status of ID4 promoter CpG island was assessed by methylation-specific PCR (MSP); ID4 mRNA level was assessed by quantitative real-time RT-PCR. Of eight cell lines, two were fully methylated, four were partially methylated, and two were not methylated. ID4 mRNA level was suppressed in fully methylated cell lines. ID4 hypermethylation was observed in 16 of 24 (67%) node-positive and seven of 36 (19%) node-negative T1 primary breast cancers matched by patient age and tumor diameter. It was a significant risk factor for nodal metastasis (OR 13.1, P ¼ 0.0004). ID4 mRNA level was suppressed in hypermethylated cancer specimens (P ¼ 0.014). ID4 may play an important suppressive role in tumor progression, and its silencing by hypermethylation may increase the risk of regional lymph node metastasis.

Inhibitor of DNA binding-4 promotes angiogenesis and growth of glioblastoma multiforme by elevating matrix GLA levels

“…ID family members including ID1, ID2, and ID3 have identified functions in tumor progression, including angiogenesis, invasion, and metastasis (Lyden et al, 1999;Benezra, 2001;Benezra et al, 2001;Folkman, 2002;Ruzinova and Benezra, 2003;, and in other circumstances ID1 and ID2 may function as tumor suppressors in rodents (Itahana et al, 2003;Sikder et al, 2003;Russell et al, 2004). The function of ID4 in tumorigenesis is similarly complex (Beger et al, 2001;Bellido et al, 2003;Chan et al, 2003;Shan et al, 2003;Umetani et al, 2004Umetani et al, , 2005 Jeon et al, 2008). ID4 seems to function as a tumor suppressor in a variety of cancers (Chan et al, 2003;Umetani et al, 2004Umetani et al, , 2005Yu et al, 2005) and is downregulated by promoter hypermethylation in several tumor types (Chan et al, 2003;Umetani et al, 2004Umetani et al, , 2005Yu et al, 2005).…”

“…The function of ID4 in tumorigenesis is similarly complex (Beger et al, 2001;Bellido et al, 2003;Chan et al, 2003;Shan et al, 2003;Umetani et al, 2004Umetani et al, , 2005 Jeon et al, 2008). ID4 seems to function as a tumor suppressor in a variety of cancers (Chan et al, 2003;Umetani et al, 2004Umetani et al, , 2005Yu et al, 2005) and is downregulated by promoter hypermethylation in several tumor types (Chan et al, 2003;Umetani et al, 2004Umetani et al, , 2005Yu et al, 2005). In sharp contrast, however, ID4 promotes some aspects of tumorigenesis in B-cell lineage ALL, glioblastoma, and sporadic breast and ovarian cancers (Beger et al, 2001;Bellido et al, 2003;Jeon et al, 2008), and its expression is heightened in bladder cancer because of gene amplification .…”

Tumoral expression of BRCA1, Estrogen receptor alpha and ID4 protein in patients with sporadic breast cancer

“…27 Although no correlation was found between the primary tumor's ID4 methylation status and its ER status, Umetani et al did not evaluate directly the relationship between ER status and ID4 mRNA levels. 32 The aim of the present study is to evaluate the relationship between tumoral expression of mRNA for BRCA1, RE and ID4 in patients with sporadic breast cancer.…”

“…Based on the genetic and epigenetic heterogeneity of primary breast cancers, and considering that metastasis occurs due to aggressive malignant clones, the authors state that finding aggressive clones with ID4 hypermethylation within a tumor lesion by MSP would be clinically useful. 32 The third protein evaluated in this study was ER alpha, that is a nuclear receptor that acts as a ligand-inducible transcription factor. ER is responsible for physiologic effects of estrogens and among other functions, participate in cell cycle and apoptosis regulation in normal mammary gland.…”

Lymphovascular invasion of colorectal cancer is correlated to SPARC expression in the tumor stromal microenvironment

“…The murine leukemia virus reverse-transcriptase (Promega, Madison, WI) with oligo-dT (GeneLink, Hawthorne, NY) priming. 47 qPCR thermo-cycling was performed in the iCycler iQ Realtime PCR Detection System (Bio-Rad Laboratories, Hercules, CA) using cDNA reverse transcribed from 250 ng of total RNA for each reaction. 48 SPARC-positive (U2OS, osteosarcoma cell line) and SPARC-negative (HT29, CRC cell line) controls and reagent controls for RT-qPCR assays were included as previously described in reference 48.…”