• Logo
  • SBMUJournals

Differential expression of aristaless-like homeobox 4: a potential marker for gastric adenocarcinoma

Fariba Ghasemvand, Navid Nezafat, Saeed Hesami Tackallou, Daruosh momenzadeh, Saeid Rahmanzadeh



Aim: Gastric cancer is the second most common cancer all over the world, approximately responsible for 3% to 10% of all cancer-related deaths. The identification of biomarkers may improve the diagnosis and treatment of gastric cancer.  Recent publications declared Aristaless-Like Homeobox 4 (ALX-4) expression levels to be up-regulated in multiple tumors.  These results were intriguing for a variety of reasons. This article explores, for the first time, examined the ALX-4 mRNA level in different grades of human Gastric adenocarcinoma comparatively,  in gastric cancer stem cell (GCSC) as well as MKN-45 cell line.

Methods: MKN-45 cell culture prepared, and gastric cancer stem cell (GCSC) isolation and identification performed by flowcytometry and then 37 fresh gastric cancer patients tissue sample were subjected for expression analysis with quantitative real-time PCR, prior to any therapeutic intervention in the comparative study for evaluation of ALX-4 gene expression.

Results: gastric cancer stem cell with cuboidal shape and positive expression for CD105, CD44, CD90 and negative for CD45, CD34 were identified. Overexpression of ALX-4 was detected in 46% (3.351±2.94, P<0.05) of gastric cancer tissue specimens and Significant high expression level of ALX-4 in GCSCs (4.31±0.04, P<0.005). The mRNA expression level of ALX-4 in MKN-45 gastric cancer cell line was 2.81±0.07 (P<0.005). ALX-4 mRNA level significantly correlation with tumor grad (P=0.004), stage (p=0.000153) and relationship with gender (P= 0.06). 

Conclusion: These sets of results documented the important role of ALX-4 in GCSCs as an indicated oncogenic role in progressive cancer and valuable target in treatment of drug resistant tumors. 


Cancer stem cell, MKN-45, ALX-4, Real-time PCR


Bozzetti F, Marubini E, Bonfanti G, et al. Subtotal versus total gastrectomy for gastric cancer: five-year survival rates in a multicenter randomized Italian trial. Italian Gastrointestinal Tumor Study Group. Ann Surg. 1999;230:170–178.

Davis P, Takeshi S. The difference in gastric cancer between Japan, USA, and Europe: what are the facts? What are the suggestions? Crit Rev Oncol Hematol. 2001;40:77–94.

Crawford J. The gastrointestinal tract. In: Robbins SLCR, Kumar V, Schoen FJ, eds. Pathologic Basis of Disease. Philadelphia: WB Saunders Co; 1994:755–783.

Schwartz G. Invasion and metastasis in gastric cancer: in vitro and in vivo models with clinical considerations. Semin Oncol. 1996;23:316– 324.

Gore R. Gastrointestinal cancer. Radiol Clin North Am. 1997;35:295– 310.

Prankin D, Whelon S, Ferlay J, et al. Cancer Incidence in Five Continents. Lyon, France: IARC Press; 1997.

Fenogilo-Preiser C, Carneiro F, Correa P, et al. Gastric carcinoma. In: Hamilton S, Aaltonin L, eds. Pathology and Genetics. Tumors of the Digestive System, vol 1. Lyon, France: Lyon Press; 2000:37–52.

Dicken BJ, Bigam DL, Cass C, Mackey JR, Joy AA, Hamilton SM. Gastric adenocarcinoma: review and considerations for future directions. Ann Surg. 2005;241(1):27-39.

Kolahdoozan S, Sadjadi A, Radmard AR, Khademi H. Five common cancers in Iran. Arch Iran Med. 2010;13(2):143-6.

Malekzadeh R, Derakhshan MH, Malekzadeh Z. Gastric cancer in Iran: epidemiology and risk factors. Arch Iran Med. 2009;12(6):576-83.

McGonnell IM, Graham A, Richardson J, Fish JL, Depew MJ, Dee CT, et al. Evolution of the Alx homeobox gene family: parallel retention and independent loss of the vertebrate Alx3 gene. Evol Dev. 2011;13(4):343-51.

Qu S, Niswender KD, Ji Q, van der Meer R, Keeney D, Magnuson MA, Wisdom R. Polydactyly and ectopic ZPA formation in Alx-4 mutant mice. Development. 1997;


Hudson R, Taniguchi-Sidle A, Boras K, Wiggan O, Hamel PA. Alx-4, a transcriptional activator whose expression is restricted to sites of epithelial-mesenchymal interactions. Dev Dyn. 1998; 213:159–69.

Chang H, Mohabir N, Done S, Hamel PA. Loss of ALX4 expression in epithelial cells and adjacent stromal cells in breast cancer. J Clin Pathol. 2009; 62:908–14.

Qu S, Tucker SC, Ehrlich JS, Levorse JM, Flaherty LA, Wisdom R, Vogt TF. Mutations in mouse Aristaless-like4 cause Strong’s luxoid polydactyly. Development. 1998;125:2711–21.

Ferrarini A, Gaillard M, Guerry F, Ramelli G, Heidi F, Keddache CV, Wieland I, Beckmann JS, Jaquemont S, www.impactjournals.com/oncotarget 13370 Oncotarget

Kariminejad A, Bozorgmehr B, Alizadeh H, Ghaderi-Sohi S, Toksoy G, Uyguner ZO, Kayserili H. Skull defects, alopecia, hypertelorism, and notched alae nasi caused by homozygous ALX4 gene mutation. Am J Med Genet A. 2014;


Ebert MP, Model F, Mooney S, Hale K, Lograsso J, Tonnes-Priddy L, Hoffmann J, Csepregi A, Röcken C, Molnar B, Schulz HU, Malfertheiner P, Lofton-Day C.

Liu WB, Han F, Du XH, Jiang X, Li YH, Liu Y, Chen HQ, Ao L, Cui ZH, Cao J, Liu JY. Epigenetic silencing of Aristaless-like homeobox-4, a potential tumor suppressor

gene associated with lung cancer. Int J Cancer. 2014; 134:1311–22.

Koso H, Tsuhako A, Lyons E, Ward JM, Rust AG, Adams DJ, Jenkins NA, Copeland NG, Watanabe S. Identification of FoxR2 as an oncogene in medulloblastoma. Cancer Res. 2014; 74:2351–61.

Shih JY, Yang PC. The EMT regulator slug and lung carcinogenesis. Carcinogenesis. 2011;32(9):1299-304.

Yuan H, Kajiyama H, Ito S, Chen D, Shibata K, Hamaguchi M, et al. HOXB13 and ALX4 induce SLUG expression for the promotion of EMT and cell invasion in ovarian cancer cells. Oncotarget. 2015;6(15):13359-70.

Gao Y, Liu T, Cheng W, Wang H. Isolation and characterization of proliferative, migratory and multidrug-resistant endometrial carcinoma-initiating cells from human type II endometrial carcinoma cell lines. Oncol Rep. 2012;28(2):527-32.

Qiao D, Zeeman AM, Deng W, Looijenga LH, Lin H. Molecular characterization of hiwi, a human member of the piwi gene family whose overexpression is correlated to seminomas. Oncogene. 2002;21(25):3988-99.

Taubert H, Greither T, Kaushal D, Wurl P, Bache M, Bartel F, et al. Expression of the stem cell self-renewal gene Hiwi and risk of tumour-related death in patients with soft-tissue sarcoma. Oncogene. 2007;26(7):1098-100.

Liu J, Ma L, Xu J, Liu C, Zhang J, Chen R, et al. Spheroid body-forming cells in the human gastric cancer cell line MKN-45 possess cancer stem cell properties. Int J Oncol. 2013;42(2):453-9.

Koso H, Tsuhako A, Lyons E, Ward JM, Rust AG, Adams DJ, et al. Identification of FoxR2 as an oncogene in medulloblastoma. Cancer Res. 2014;74(8):2351-61.

Martinet D. Potocki-Shaffer deletion encompassing ALX4 in a patient with frontonasal dysplasia phenotype. Am J Med Genet A. 2014; 164A:346–52.

Aristaless-like homeobox-4 gene methylation is a potential marker for colorectal adenocarcinomas. Gastroenterology. 2006; 131:1418–30.

Ushiku T, Shinozaki A, Shibahara J, Iwasaki Y, Tateishi Y, Funata N, et al. SALL4 represents fetal gut differentiation of gastric cancer, and is diagnostically useful in distinguishing hepatoid gastric carcinoma from hepatocellular carcinoma. Am J Surg Pathol. 2010;34(4):533-40.

DOI: https://doi.org/10.22037/ghfbb.v0i0.865