Analysis of SEMA3B methylation and expression patterns in gastric cancer tissue and cell lines

Chen,Renpin; Zhuge,Xiaoju; Huang,Zhiming; Lu,Deyi; Ye,Xiaohua; Chen,Chao; Yu,Jieyu; Lu,Guangrong

doi:10.3892/or.2014.2972

Analysis of SEMA3B methylation and expression patterns in gastric cancer tissue and cell lines

Authors:
- Renpin Chen
- Xiaoju Zhuge
- Zhiming Huang
- Deyi Lu
- Xiaohua Ye
- Chao Chen
- Jieyu Yu
- Guangrong Lu
View Affiliations

Affiliations: Department of Gastroenterology and Hepatology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325035, P.R. China, Department of Gastroenterology, Jinhua Municipal Central Hospital, Jinhua, Zhejiang 32100, P.R. China, Department of Gastroenterology and Hepatology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China
Published online on: January 9, 2014 https://doi.org/10.3892/or.2014.2972
Pages: 1211-1218

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The family of semaphorins has been demonstrated to possess tumor suppressor activity, in which semaphorin 3B (SEMA3B) is differentially expressed in several types of tumors. The relationship between SEMA3B expression and its clinical significance in gastric cancer (GC) is currently unclear. In the present study, the expression and methylation status of the SEMA3B gene were detected by quantitative PCR and bisulfite sequencing PCR (BSP). Data indicated that the levels of SEMA3B mRNA decreased in gastric tumor tissues and the methylation status of SEMA3B in the tumor group was higher than the paired normal tissues. By BSP, the SEMA3B gene showed high methylated status which was detected in all 4 cell lines (AGS, BGC-823, MGC-803 and SGC-7901). Treatment of the cells with 5-Aza-2'-deoxycytidine revealed clearly elevated mRNA levels of SEMA3B. These results were further confirmed by western blot analysis of Sema3b protein expression. At the same time, increased expression of p53 mRNA in BGC-823, MGC-803 was detected and indicated that p53 may be involved in the regulation of SEMA3B expression in specific genetic background. Downregulation of SEMA3B was negatively correlated with tumor size and N staging in GC (p<0.05). In conclusion, CpG methylation of SEMA3B epigenetically regulates SEMA3B expression during development of GC. Furthermore, 5-Aza-2'-deoxycytidine could reverse the hypermethylation status of SEMA3B, which may benefit future studies exploring the application of demethylating agents in clinical usage of GC.

View Figures

View References

1	Parkin DM, Bray F, Ferlay J and Pisani P: Global cancer statistics, 2002. CA Cancer J Clin. 55:74–108. 2005. View Article : Google Scholar
2	Rivera F, Vega-Villegas ME and Lopez-Brea MF: Chemotherapy of advanced gastric cancer. Cancer Treat Rev. 33:315–324. 2007. View Article : Google Scholar
3	Yazdani U and Terman JR: The semaphorins. Genome Biol. 7:2112006. View Article : Google Scholar : PubMed/NCBI
4	Chilton JK and Guthrie S: Cranial expression of class 3 secreted semaphorins and their neuropilin receptors. Dev Dyn. 228:726–733. 2003. View Article : Google Scholar : PubMed/NCBI
5	Giger RJ, Cloutier JF, Sahay A, et al: Neuropilin-2 is required in vivo for selective axon guidance responses to secreted semaphorins. Neuron. 25:29–41. 2000. View Article : Google Scholar : PubMed/NCBI
6	Falk J, Bechara A, Fiore R, et al: Dual functional activity of semaphorin 3B is required for positioning the anterior commissure. Neuron. 48:63–75. 2005. View Article : Google Scholar : PubMed/NCBI
7	Nawabi H, Briancon-Marjollet A, Clark C, et al: A midline switch of receptor processing regulates commissural axon guidance in vertebrates. Genes Dev. 24:396–410. 2010. View Article : Google Scholar : PubMed/NCBI
8	Tamagnone L and Comoglio PM: To move or not to move? Semaphorin signalling in cell migration. EMBO Rep. 5:356–361. 2004. View Article : Google Scholar : PubMed/NCBI
9	Sekido Y, Bader S, Latif F, et al: Human semaphorins A(V) and IV reside in the 3p21.3 small cell lung cancer deletion region and demonstrate distinct expression patterns. Proc Natl Acad Sci USA. 93:4120–4125. 1996. View Article : Google Scholar : PubMed/NCBI
10	Tomizawa Y, Sekido Y, Kondo M, et al: Inhibition of lung cancer cell growth and induction of apoptosis after reexpression of 3p21.3 candidate tumor suppressor gene SEMA3B. Proc Natl Acad Sci USA. 98:13954–13959. 2001. View Article : Google Scholar : PubMed/NCBI
11	Tse C, Xiang RH, Bracht T and Naylor SL: Human Semaphorin 3B (SEMA3B) located at chromosome 3p21.3 suppresses tumor formation in an adenocarcinoma cell line. Cancer Res. 62:542–546. 2002.PubMed/NCBI
12	Tischoff I, Markwarth A, Witzigmann H, et al: Allele loss and epigenetic inactivation of 3p21.3 in malignant liver tumors. Int J Cancer. 115:684–689. 2005. View Article : Google Scholar
13	Ochi K, Mori T, Toyama Y, Nakamura Y and Arakawa H: Identification of semaphorin3B as a direct target of p53. Neoplasia. 4:82–87. 2002. View Article : Google Scholar : PubMed/NCBI
14	Castro-Rivera E, Ran S, Brekken RA and Minna JD: Semaphorin 3B inhibits the phosphatidylinositol 3-kinase/Akt pathway through neuropilin-1 in lung and breast cancer cells. Cancer Res. 68:8295–8303. 2008. View Article : Google Scholar : PubMed/NCBI
15	Castro-Rivera E, Ran S, Thorpe P and Minna JD: Semaphorin 3B (SEMA3B) induces apoptosis in lung and breast cancer, whereas VEGF165 antagonizes this effect. Proc Natl Acad Sci USA. 101:11432–11437. 2004. View Article : Google Scholar : PubMed/NCBI
16	Koyama N, Zhang J, Huqun, et al: Identification of IGFBP-6 as an effector of the tumor suppressor activity of SEMA3B. Oncogene. 27:6581–6589. 2008. View Article : Google Scholar : PubMed/NCBI
17	Varshavsky A, Kessler O, Abramovitch S, et al: Semaphorin-3B is an angiogenesis inhibitor that is inactivated by furin-like pro-protein convertases. Cancer Res. 68:6922–6931. 2008. View Article : Google Scholar : PubMed/NCBI
18	Rolny C, Capparuccia L, Casazza A, et al: The tumor suppressor semaphorin 3B triggers a prometastatic program mediated by interleukin 8 and the tumor microenvironment. J Exp Med. 205:1155–1171. 2008. View Article : Google Scholar : PubMed/NCBI
19	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001.
20	Vailaya A, Bluvas P, Kincaid R, Kuchinsky A, Creech M and Adler A: An architecture for biological information extraction and representation. Bioinformatics. 21:430–438. 2005. View Article : Google Scholar : PubMed/NCBI
21	Li LC and Dahiya R: MethPrimer: designing primers for methylation PCRs. Bioinformatics. 18:1427–1431. 2002. View Article : Google Scholar : PubMed/NCBI
22	Gluzman-Poltorak Z, Cohen T, Herzog Y and Neufeld G: Neuropilin-2 is a receptor for the vascular endothelial growth factor (VEGF) forms VEGF-145 and VEGF-165. J Biol Chem. 275:299222000. View Article : Google Scholar : PubMed/NCBI
23	Soker S, Miao HQ, Nomi M, Takashima S and Klagsbrun M: VEGF165 mediates formation of complexes containing VEGFR-2 and neuropilin-1 that enhance VEGF165-receptor binding. J Cell Biochem. 85:357–368. 2002. View Article : Google Scholar : PubMed/NCBI
24	Haiko P, Makinen T, Keskitalo S, et al: Deletion of vascular endothelial growth factor C (VEGF-C) and VEGF-D is not equivalent to VEGF receptor 3 deletion in mouse embryos. Mol Cell Biol. 28:4843–4850. 2008. View Article : Google Scholar : PubMed/NCBI
25	Li K, Chen MK, Li LY, et al: The predictive value of semaphorins 3 expression in biopsies for biochemical recurrence of patients with low- and intermediate-risk prostate cancer. Neoplasma. 60:683–689. 2013. View Article : Google Scholar : PubMed/NCBI
26	Pronina IV, Loginov VI, Prasolov VS, et al: Alteration of SEMA3B gene expression levels in epithelial tumors. Mol Biol (Mosk). 43:439–445. 2009.(In Russian).
27	Riquelme E, Tang M, Baez S, et al: Frequent epigenetic inactivation of chromosome 3p candidate tumor suppressor genes in gallbladder carcinoma. Cancer Lett. 250:100–106. 2007. View Article : Google Scholar : PubMed/NCBI
28	Kuroki T, Trapasso F, Yendamuri S, et al: Allelic loss on chromosome 3p21.3 and promoter hypermethylation of semaphorin 3B in non-small cell lung cancer. Cancer Res. 63:3352–3355. 2003.PubMed/NCBI
29	Hesson L, Bieche I, Krex D, et al: Frequent epigenetic inactivation of RASSF1A and BLU genes located within the critical 3p21.3 region in gliomas. Oncogene. 23:2408–2419. 2004.PubMed/NCBI
30	Bernal C, Vargas M, Ossandon F, et al: DNA methylation profile in diffuse type gastric cancer: evidence for hypermethylation of the BRCA1 promoter region in early-onset gastric carcinogenesis. Biol Res. 41:303–315. 2008. View Article : Google Scholar : PubMed/NCBI
31	Bernal C, Aguayo F, Villarroel C, et al: Reprimo as a potential biomarker for early detection in gastric cancer. Clin Cancer Res. 14:6264–6269. 2008. View Article : Google Scholar : PubMed/NCBI
32	Zhou X, Ma L, Li J, Gu J, Shi Q and Yu R: Effects of SEMA3G on migration and invasion of glioma cells. Oncol Rep. 28:269–275. 2012.PubMed/NCBI