Methylation pattern of ALX4 gene promoter as a potential biomarker for blood-based early detection of colorectal cancer

Abstract

Background:To develop a non-invasive screening method for colorectal cancer, we evaluated the methylation of ALX4 gene promoter in serum samples from patients with colorectal cancer (CRC) and equal number of healthy individuals.Materials and Methods:In serum samples from 25 patients with colorectal cancer and 25 healthy control subjects, isolated serum free-floating DNA was treated with sodium bisulfite and analyzed by methylation-specific polymerase chain reaction (MSP) with primers specific for methylated or unmethylated promoter CpG island sequences of the ALX4 gene.Results:Methylation of the ALX4 gene promoter was present in the serum DNA of patients with adenoma and colorectal cancer. A sensitivity of 68% and specificity of 88% were achieved in the detection of promoter methylation in colorectal neoplasia samples. The difference in methylation status of the ALX4 promoter between the patients with colorectal neoplasia and the control group was statistically highly significant (P < 0.001).Conclusions:The results indicate that this serum free DNA test of methylation of the ALX4 gene promoter is a sensitive and specific method. Therefore in combination with other useful markers it seems ALX4 has the potential of a clinically useful test for the early detection of colorectal cancer.

Methylation as a critical epigenetic process during tumor progressions among Iranian population: an overview

“…Homeobox protein aristaless-like (ALX4) is a homeodomain transcription factor associated with bone, skin, and hair follicle development [ 182 , 183 ]. It has been reported that there was a significant difference of ALX4 methylation status between a sample of Iranian colorectal cancer patients and controls which introduced that as an efficient marker for the early detection of colorectal cancer in this population [ 62 ]. Similarly, ALX4 methylation was observed among German patients with colorectal, esophageal, and gastric cancers [ 184 ].…”

“… Yari [ 61 ] developmental factors ALX4 2015 Colorectal 25 patients 25 controls Serum Hyper methylation. Salehi [ 62 ] PAX5 2018 Gastric 35 patients 35 controls Blood Hyper methylation. Haghverdi [ 63 ] MiR-129-2 2019 Gastric 50 N/T Tissue Hyper methylation.…”

Epigenetic Modifications as Biomarkers of Tumor Development, Therapy Response, and Recurrence across the Cancer Care Continuum

“…Tests like Epi proColon show great clinical potential because patients already prefer these non-invasive tests over colonoscopy/sigmoidoscopy for screening, and SEPT9 methylation is superior at detecting early stage disease over other non-invasive tests like fecal occult blood testing [ 100 , 101 , 102 ]. Combinations of other hypermethylated genes as stool or blood biomarkers have not displayed the high sensitivity observed in the SEPT9 methylation assays [ 89 , 103 ].…”

Blood-based DNA Methylation Biomarkers for Early Detection of Colorectal Cancer

“…Aberrant CpG island methylation of the promoter region of p16 and IGFBP7 bypasses this dormant state and drives MVHPs further to sessile serrated adenomas (SSAs) [ 97 ]. In blood samples, hypermethylated ALX4 [ 80 , 98 ], NEUROG1 [ 99 ], APC [ 100 ], 6-O-Methylguanine-DNA Methyltransferase (MGMT) [ 101 ], MLH1 [ 82 ], HLTF [ 102 ], Ras association domain family member 2 (RASSF2A) [ 101 ], Syndecan 2 (SDC2) [ 103 ], SEPT9, Preprotachykinin-1 (TAC1) [ 104 ] and WIF1 [ 101 ] were detected in early stage CRC; hypermethylated HPP1, HLTF, secreted Frizzled Related Protein 2 (SFRP2) [ 105 ], VIM [ 106 ], tissue factor pathway inhibitor 2 (TFPI2) [ 107 , 108 ] and p16 [ 89 ] were found positively correlated with distant tumor metastasis; hypermethylated ALX4, fibrillin-2 (FBN2), HPP1 (Alias TMEFF2) [ 102 ], HLTF [ 83 ], p16 [ 91 ], TMEFF1 [ 102 ] and VIM [ 99 ] were associated with poor prognosis; and hypermethylated HLTF [ 109 ], HPP1 [ 109 ], runt- related transcription factor 3 (RUNX3) [ 110 ], p16 [ 91 ] and TFPI2 [ 107 , 108 ] were associated with CRC recurrence. It is conceivable that a robust biomarker panel of methylated genes will be developed into a clinically accurate CRC screening method in the future, and the development of blood-based biomarkers should improve patient compliance and the detection of CRC at early stage.…”