Status of integrin subunit alpha 4 promoter DNA methylation in colorectal cancer and other malignant tumors: a systematic review and meta-analysis

Sima Jafarpour , Maryam Yazdi, Reza Nedaeinia, Nasimeh Vatandoost, Gordon A. Ferns, Rasoul Salehi


Background and purpose: Although many recent studies have analyzed the validation of integrin subunit alpha 4 (ITGA4) biomarker for cancer detection in patients with various malignancies, the diagnostic value of ITGA4 methylation for malignant tumors remains uncertain. We performed a systematic review and meta-analysis to unravel the relationship between ITGA4 promoter methylation status and malignant tumors.

Experimental approach: A meta-analysis was performed using the metaphor package in R 3.5 and Meta-Disc 1.4 software. Data were derived from a search of main electronic databases up to January 2022. SROC analysis was used to evaluate the status of ITGA4 promoter methylation in colorectal cancer (CRC) and other cancers. A total of 1232 tumor samples and 649 non-tumor samples from 13 studies were analyzed.

Findings/Results: The pooled results including all types of cancer provided evidence that ITGA4 hypermethylation was more frequent in tumor samples than non-tumor samples (OR 13.32, 95% CI 7.96-22.29). Methylation of ITGA4 has a pooled sensitivity of 0.95 (95% CI: 0.94-0.97), a pooled specificity of 0.57 (95% CI: 0.54-0.60), and an area under the curve (AUC) of 0.94. When the analysis was performed independently for CRC, it revealed a higher association (OR = 20.77, 95% CI: 9.15-47.15). The assessment of ITGA4 methylation of tissue samples resulted in a pooled sensitivity of 0.99 (95% CI: 0.97-1.00) and a pooled specificity of 0.90 (95% CI: 0.86-0.93), and AUC of 0.94 for the diagnosis of CRC.

Conclusion and implications: ITGA4 methylation analysis is a reliable method for CRC screening in tissue samples.


Colorectal cancer; ITGA4 gene; Meta-analysis; Promoter methylation.

Full Text:



Siegel RL, Miller KD, Jemal A. Cancer statistics, 2020. CA Cancer J Clin. 2020;70(1):7-30.DOI: 10.3322/caac.21590.

Tavangar S, Bohlooli S, Razzaghi-Asl N. Synthesis and cytotoxic effect of a few N-heteroaryl enamino amides and dihydropyrimidinethiones on AGS and MCF-7 human cancer cell lines. Res Pharm Sci. 2020;15(2):154-163.DOI: 10.4103/1735-5362.283815.

Allemani C, Weir HK, Carreira H, Harewood R, Spika D, Wang XS, et al. Global surveillance of cancer survival 1995-2009: analysis of individual data for 25,676,887 patients from 279 population-based registries in 67 countries (CONCORD-2). Lancet. 2015;385(9972):977-1010.DOI: 10.1016/s0140-6736(14)62038-9.

Fitzmaurice C, Dicker D, Pain A, Hamavid H, Moradi-Lakeh M, MacIntyre MF, et al. The global burden of cancer 2013. JAMA Oncol. 2015;1(4):505-527.DOI: 10.1001/jamaoncol.2015.0735.

Schiffman JD, Fisher PG, Gibbs P. Early detection of cancer: past, present, and future. Am Soc Clin Oncol Educ Book. 2015:57-65.DOI: 10.14694/EdBook_AM.2015.35.57.

Ghavifekr Fakhr M, Farshdousti Hagh M, Shanehbandi D, Baradaran B. DNA methylation pattern as important epigenetic criterion in cancer. Genet Res Int. 2013;2013:317569.DOI: 10.1155/2013/317569.

Pfeifer GP. Defining driver DNA methylation changes in human cancer. Int J Mol Sci. 2018;19(4),1-13.DOI: 10.3390/ijms19041166.

Daura-Oller E, Cabre M, Montero MA, Paternain JL, Romeu A. Specific gene hypomethylation and cancer: new insights into coding region feature trends. Bioinformation. 2009;3(8):340-343.DOI: 10.6026/97320630003340.

Bardhan K, Liu K. Epigenetics and colorectal cancer pathogenesis. Cancers (Basel). 2013;5(2):676-713.DOI: 10.3390/cancers5020676.

Vatandoost N, Ghanbari J, Mojaver M, Avan A, Ghayour-Mobarhan M, Nedaeinia R, et al. Early detection of colorectal cancer: from conventional methods to novel biomarkers. J Cancer Res Clin Oncol. 2016;142(2):341-351.DOI: 10.1007/s00432-015-1928-z.

Locke WJ, Guanzon D, Ma C, Liew YJ, Duesing KR, Fung KYC, et al. DNA methylation cancer biomarkers: translation to the clinic. Front Genet. 2019;10:1150,1-44.DOI: 10.3389/fgene.2019.01150.

Bakhtiari H, Palizban AA, Khanahmad H, Mofid MR. Aptamer-based approaches for in vitro molecular detection of cancer. Res Pharm Sci. 2020;15(2): 107-122.DOI: 10.4103/1735-5362.283811.

Sajjadi RS, Modarressi MH, Tabatabaiefar MA. JPX and LINC00641 ncRNAs expression in prostate tissue: a case-control study. Res Pharm Sci. 2021;16(5):493-504.DOI: 10.4103/1735-5362.323916.

Laugsand EA, Brenne SS, Skorpen F. DNA methylation markers detected in blood, stool, urine, and tissue in colorectal cancer: a systematic review of paired samples. Int J Colorectal Dis. 2021;36(2): 239-251.DOI: 10.1007/s00384-020-03757-x.

Attia HR, Ibrahim MH, El-Aziz SHA, Hassan NM, Osman RA, Hagag HA, et al. ITGA4 gene methylation status in chronic lymphocytic leukemia. Future Sci OA. 2020;6(7):Fso583,1-12.DOI: 10.2144/fsoa-2020-0034.

Zhang W, Shang S, Yang Y, Lu P, Wang T, Cui X, et al. Identification of DNA methylation-driven genes by integrative analysis of DNA methylation and transcriptome data in pancreatic adenocarcinoma. Exp Ther Med. 2020;19(4):2963-2972.DOI: 10.3892/etm.2020.8554.

Darzi L, Boshtam M, Shariati L, Kouhpayeh S, Gheibi A, Mirian M, et al. The silencing effect of miR-30a on ITGA4 gene expression in vitro: an approach for gene therapy. Res Pharm Sci. 2017;12(6):456-464.DOI: 10.4103/1735-5362.217426.

Arnaout MA, Goodman SL, Xiong JP. Structure and mechanics of integrin-based cell adhesion. Curr opin in cell biol. 2007;19(5):495-507.DOI: 10.1016/

Huttenlocher A, Horwitz AR. Integrins in cell migration. Cold Spring Harb Perspect Biol. 2011;3(9):a005074,1-17.DOI: 10.1101/cshperspect.a005074.

Pulkka OP, Mpindi JP, Tynninen O, Nilsson B, Kallioniemi O, Sihto H, et al. Clinical relevance of integrin alpha 4 in gastrointestinal stromal tumours. J Cell Mol Med. 2018;22(4):2220-2230.DOI: 10.1111/jcmm.13502.

Deb M, Sengupta D, Patra SK. Integrin-epigenetics: a system with imperative impact on cancer. Cancer Metastasis Rev. 2012;31(1-2):221-234.DOI: 10.1007/s10555-011-9341-9.

Klemke M, Weschenfelder T, Konstandin MH, Samstag Y. High affinity interaction of integrin alpha4beta1 (VLA-4) and vascular cell adhesion molecule 1 (VCAM-1) enhances migration of human melanoma cells across activated endothelial cell layers. J Cell Physiol. 2007;212(2):368-374.DOI: 10.1002/jcp.21029.

Strelnikov VV, Kuznetsova EB, Tanas AS, Rudenko VV, Kalinkin AI, Poddubskaya EV, et al. Abnormal promoter DNA hypermethylation of the integrin, nidogen, and dystroglycan genes in breast cancer. Sci Rep. 2021;11(1):2264.DOI: 10.1038/s41598-021-81851-y.

Nurzat Y, Su W, Min P, Li K, Xu H, Zhang Y. Identification of therapeutic targets and prognostic biomarkers among integrin subunits in the skin cutaneous melanoma microenvironment. Front Oncol. 2021;11:751875,1-16.DOI: 10.3389/fonc.2021.751875.

Mo J, Zhang J, Huang H, Liu C, Cheng Y, Mo Y, et al. The early predictive effect of low expression of the ITGA4 in colorectal cancer. J Gastrointest Oncol. 2022;13(1):265-278.DOI: 10.21037/jgo-22-92.

Young SA, McCabe KE, Bartakova A, Delaney J, Pizzo DP, Newbury RO, et al. Integrin α4 enhances metastasis and may be associated with poor prognosis in MYCN-low neuroblastoma. PLoS One. 2015;10(5):e0120815.DOI: 10.1371/journal.pone.0120815,1-17.

Gerecke C, Scholtka B, Löwenstein Y, Fait I, Gottschalk U, Rogoll D, et al. Hypermethylation of ITGA4, TFPI2 and VIMENTIN promoters is increased in inflamed colon tissue: putative risk markers for colitis-associated cancer. J Cancer Res Clin Oncol. 2015;141(12):2097-2107.DOI: 10.1007/s00432-015-1972-8.

Morandi L, Gissi D, Tarsitano A, Asioli S, Gabusi A, Marchetti C, et al. CpG location and methylation level are crucial factors for the early detection of oral squamous cell carcinoma in brushing samples using bisulfite sequencing of a 13-gene panel. Clin Epigenetics. 2017;9:85,1-17.DOI: 10.1186/s13148-017-0386-7.

Liu X, Wen J, Li C, Wang H, Wang J, Zou H. High-yield methylation markers for stool-based detection of colorectal cancer. Dig Dis Sci. 2020;65(6):1710-1719.DOI: 10.1007/s10620-019-05908-9.

Zhang Y, Wu Q, Xu L, Wang H, Liu X, Li S, et al. Sensitive detection of colorectal cancer in peripheral blood by a novel methylation assay. Clin Epigenetics. 2021;13(1):90,1-10.DOI: 10.1186/s13148-021-01076-8.

Moher D, Liberati A, Tetzlaff J, Altman DG, Group P. Preferred reporting items for systematic reviews and meta-analyses: the PRISMA statement. Ann Intern Med. 2009;151(4):264-269,W64.DOI: 10.7326/0003-4819-151-4-200908180-00135.

Jackson D, Law M, Stijnen T, Viechtbauer W, White IR. A comparison of seven random-effects models for meta-analyses that estimate the summary odds ratio. Stat Med. 2018;37(7):1059-1085.DOI: 10.1002/sim.7588.

Bradburn MJ, Deeks JJ, Berlin JA, Russell Localio A. Much ado about nothing: a comparison of the performance of meta-analytical methods with rare events. Stat Med. 2007;26(1):53-77.DOI: 10.1002/sim.2528.

Higgins JP, Thompson SG, Deeks JJ, Altman DG. Measuring inconsistency in meta-analyses. BMJ.2003;327(7414):557-560.DOI: 10.1136/bmj.327.7414.557.

Jones CM, Athanasiou T. Summary receiver operating characteristic curve analysis techniques in the evaluation of diagnostic tests. Ann Thorac Surg. 2005;79(1):16-20.DOI: 10.1016/j.athoracsur.2004.09.040.

Deeks JJ, Macaskill P, Irwig L. The performance of tests of publication bias and other sample size effects in systematic reviews of diagnostic test accuracy was assessed. J Clin Epidemiol. 2005;58(9):882-893.DOI: 10.1016/j.jclinepi.2005.01.016.

ViechtbauerV. Conducting meta-analyses in R with the Metafor package. J Stati Soft. 2010;36(3):1-48.DOI: 10.18637/jss.v036.i03

Yu J, Zhu T, Wang Z, Zhang H, Qian Z, Xu H, et al. A novel set of DNA methylation markers in urine sediments for sensitive/specific detection of bladder cancer. Clin Cancer Res . 2007;13(24):7296-7304.DOI: 10.1158/1078-0432.ccr-07-0861.

Kurdyukov S, Bullock M. DNA methylation analysis: choosing the right method. Biology. 2016; 5(1):3,1-21. DOI: 10.3390/biology5010003.

Asl MM, Asl JM, Naghitorabi M. Comparison of the effects of olsalazine and decitabine on the expression of CDH1 and uPA genes and cytotoxicity in MDA-MB-231 breast cancer cells. Res Pharm Sci. 2021;16(3):278-285.DOI: 10.4103/1735-5362.314826.

Kondo Y. Epigenetic cross-talk between DNA methylation and histone modifications in human cancers. Yonsei Med J. 2009;50(4):455-463.DOI: 10.3349/ymj.2009.50.4.455.

Naghitorabi M, Mohammadi Asl J, Mir Mohammad Sadeghi H, Rabbani M, Jafarian-Dehkordi A, Javanmard HS. Quantitative evaluation of DNMT3B promoter methylation in breast cancer patients using differential high resolution melting analysis. Res Pharm Sci. 2013;8(3):167-175 PMID: 24019826.

Skvortsova K, Stirzaker C, Taberlay P. The DNA methylation landscape in cancer. Essays Biochem. 2019;63(6):797-811.DOI: 10.1042/ebc20190037.

Campbell ID, Humphries MJ. Integrin structure, activation, and interactions. Cold Spring Harb Perspect Biol. 2011;3(3).DOI: 10.1101/cshperspect.a004994.

Strelnikov VV, Kuznetsova EB, Tanas AS, Rudenko VV, Kalinkin AI, Poddubskaya EV, et al. Abnormal promoter DNA hypermethylation of the integrin, nidogen, and dystroglycan genes in breast cancer. Sci Rep. 2021;11(1):2264,1-14.DOI: 10.1038/s41598-021-81851-y.

Wu J, Gu Y, Xiao Y, Xia C, Li H, Kang Y, et al. Characterization of DNA methylation associated gene regulatory networks during stomach cancer progression. Front Genet. 2019;9:711,1-9.DOI: 10.3389/fgene.2018.00711.

Chang E, Park DI, Kim YJ, Kim BK, Park JH, Kim HJ, et al. Detection of colorectal neoplasm using promoter methylation of ITGA4, SFRP2, and p16 in stool samples: a preliminary report in Korean patients. Hepatogastroenterology. 2010;57(101):720-727.PMID: 21033217.

Uhm KO, Lee JO, Lee YM, Lee ES, Kim HS, Park SH. Aberrant DNA methylation of integrin alpha4: a potential novel role for metastasis of cholangiocarcinoma. J Cancer Res Clin Oncol. 2010;136(2):187-194.DOI: 10.1007/s00432-009-0646-9.

Park J, Song SH, Kim TY, Choi MC, Jong HS, Kim TY, et al. Aberrant methylation of integrin α4 gene in human gastric cancer cells. Oncogene. 2004;23(19):3474-3480.DOI: 10.1038/sj.onc.1207470.

Dong SM, Lee EJ, Jeon ES, Park CK, Kim KM. Progressive methylation during the serrated neoplasia pathway of the colorectum. Mod Pathol. 2005;18(2):170-178.DOI: 10.1038/modpathol.3800261.

Ausch C, Kim YH, Tsuchiya KD, Dzieciatkowski S, Washington MK, Paraskeva C, et al. Comparative analysis of PCR-based biomarker assay methods for colorectal polyp detection from fecal DNA. Clin Chem. 2009;55(8):1559-1563.DOI: 10.1373/clinchem.2008.122937.


  • There are currently no refbacks.

Creative Commons LicenseThis work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.