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MiR-6872 host gene SEMA3B and its antisense lncRNA SEMA3B-AS1 function synergistically to suppress gastric cardia adenocarcinoma progression

  • Wei Guo
  • Xiaoliang Liang
  • Lei Liu
  • Yanli Guo
  • Supeng Shen
  • Jia Liang
  • Zhiming DongEmail author
Original Article
  • 132 Downloads

Abstract

Background

Semaphorin 3B (SEMA3B) is frequently inactivated in several carcinomas. However, as the host gene of miR-6872, the roles of SEMA3B, antisense lncRNA SEMA3B-AS1, and miR-6872 in gastric cardia adenocarcinoma (GCA) tumorigenesis have not been clarified.

Methods

The expression levels of SEMA3B, SEMA3B-AS1, and miR-6872 were respectively detected by qRT-PCR, western blot, or immunohistochemical staining assays. The methylation status was determined by BGS and BS-MSP methods. In vitro assays were preformed to explore the biological effects of SEMA3B, SEMA3B-AS1, and miR-6872-5p in gastric cancer cells. Chromatin immunoprecipitation assay was used to detect the binding of protein to DNA. The interaction of SEMA3B-AS1 with MLL4 was identified by RNA immunoprecipitation and RNA pull-down assays.

Results

Frequent downregulation of SEMA3B, SEMA3B-AS1, and miR-6872 was detected in GCA tissues and gastric cancer cells. Aberrant hypermethylation of the promoter region was more tumor specific and was negatively correlated with the expression level of SEMA3B, SEMA3B-AS1, and miR-6872-5p. Transcription factor Sp1 activated SEMA3B or SEMA3B-AS1 transcription and CpG sites hypermethylation within promoter region eliminated Sp1 binding ability. Overexpression of SEMA3B and SEMA3B-AS1 inhibited gastric cancer cell proliferation, migration, and invasion in vitro. SEMA3B-AS1 induced the expression of SEMA3B by interacting with MLL4. ZNF143 might be the target gene of miR-6872-5p and miR-6872-5p functioning synergistically with SEMA3B to suppress cell invasion. Furthermore, SEMA3B, SEMA3B-AS1, and miR-6872-5p expression levels were associated with GCA patients’ survival.

Conclusions

SEMA3B, SEMA3B-AS1, and miR-6872 may act as tumor suppressors and may serve as potential targets for antitumor therapy.

Keywords

SEMA3B SEMA3B-AS1 MiR-6872 Gastric cardia adenocarcinoma Expression 

Notes

Acknowledgements

This study was supported by Grants from the National Natural Science Foundation (Nos. 81472335, 81572441, 81772612), Natural Science Foundation of Hebei Province (Nos. H2015206196 and H2015206420).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

The study was approved by the Ethics Committee of the Fourth Hospital, Hebei Medical University.

Informed consent

Informed consent was obtained from all participants included in the study.

Supplementary material

10120_2019_924_MOESM1_ESM.docx (1 mb)
Supplementary material 1 (DOCX 1036 KB)

References

  1. 1.
    Vial M, Grande L, Pera M. Epidemiology of adenocarcinoma of the esophagus, gastric cardia, and uppergastric third. Recent Results Cancer Res. 2010;182:1–17.Google Scholar
  2. 2.
    Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65:87–108.CrossRefGoogle Scholar
  3. 3.
    Chow WH, Finkle WD, McLaughlin JK, Frankl H, Ziel HK, Fraumeni JF. The relation of gastroesophageal reflux disease and its treatment to adenocarcinomas of the esophagus and gastric cardia. JAMA. 1995;274:474–7.CrossRefGoogle Scholar
  4. 4.
    Wang LD, Zheng S, Zheng ZY, Casson AG. Primary adenocarcinomas of lower esophagus, esophagogastric junction and gastric cardia: in special reference to China. World J Gastroenterol. 2003;9:1156–64.CrossRefGoogle Scholar
  5. 5.
    Plummer M, Franceschi S, Vignat J, Forman D, de Martel C. Global burden of gastric cancer attributable to Helicobacter pylori. Int J Cancer. 2015;136:487–90.CrossRefGoogle Scholar
  6. 6.
    Wistuba II, Behrens C, Virmani AK, Mele G, Milchgrub S, Girard L, et al. High resolution chromosome 3p allelotyping of human lung cancer and preneoplastic/preinvasive bronchial epithelium reveals multiple, discontinuous sites of 3p allele loss and three regions of frequent breakpoints. Cancer Res. 2000;60:1949–60.Google Scholar
  7. 7.
    Lerman MI, Minna JD. The 630-kb lung cancer homozygous deletion region on human chromosome 3p21.3: identification and evaluation of the resident candidate tumor suppressor genes. The international lung cancer chromosome 3p21.3 Tumor Suppressor Gene Consortium. Cancer Res. 2000;60:6116–33.Google Scholar
  8. 8.
    Chen J, Brevet A, Blanquet S, Plateau P. Control of 5′,5′-dinucleoside triphosphate catabolism by APH1, a Saccharomyces cerevisiae analog of human FHIT. J Bacteriol. 1998;180:2345–9.Google Scholar
  9. 9.
    Yue W, Dacic S, Sun Q, Landreneau R, Guo M, Zhou W, et al. Frequent inactivation of RAMP2, EFEMP1 and Dutt1 in lung cancer by promoter hypermethylation. Clin Cancer Res. 2007;13:4336–44.CrossRefGoogle Scholar
  10. 10.
    Guo W, Dong Z, Chen Z, Yang Z, Wen D, Kuang G, et al. Aberrant CpG Island hypermethylation of RASSF1A in gastric cardia adenocarcinoma. Cancer Invest. 2009;27:459–65.CrossRefGoogle Scholar
  11. 11.
    Loginov VI, Dmitriev AA, Senchenko VN, Pronina IV, Khodyrev DS, Kudryavtseva AV, et al. Tumor suppressor function of the SEMA3B Gene in human lung and renal cancers. PLoS One. 2015;10:e0123369.CrossRefGoogle Scholar
  12. 12.
    Pang CH, Du W, Long J, Song LJ. Mechanism of SEMA3B gene silencing and clinical significance in glioma. Genet Mol Res. 2016.  https://doi.org/10.4238/gmr.15017664.Google Scholar
  13. 13.
    Castro-Rivera E, Ran S, Brekken RA, Minna JD. Semaphorin 3B inhibits the phosphatidylinositol 3-kinase/Akt pathway through neuropilin-1 in lung and breast cancer cells. Cancer Res. 2008;68:8295–303.CrossRefGoogle Scholar
  14. 14.
    Chen R, Zhuge X, Huang Z, Lu D, Ye X, Chen C, et al. Analysis of SEMA3B methylation and expression patterns in gastric cancer tissue and cell lines. Oncol Rep. 2014;31:1211–8.CrossRefGoogle Scholar
  15. 15.
    Riquelme E, Tang M, Baez S, Diaz A, Pruyas M, Wistuba II, et al. Frequent epigenetic inactivation of chromosome 3p candidate tumor suppressor genes in gallbladder carcinoma. Cancer Lett. 2007;250:100–6.CrossRefGoogle Scholar
  16. 16.
    Luo Y, Raible D, Raper JA. Collapsin: a protein in brain that induces the collapse and paralysis of neuronal growth cones. Cell. 1993;75:217–27.CrossRefGoogle Scholar
  17. 17.
    Gaur P, Bielenberg DR, Samuel S, Bose D, Zhou Y, Gray MJ, et al. Role of class 3 semaphorins and their receptors in tumor growth and angiogenesis. Clin Cancer Res. 2009;15:6763–70.CrossRefGoogle Scholar
  18. 18.
    Grote HJ, Schmiemann V, Geddert H, Rohr UP, Kappes R, Gabbert HE, et al. Aberrant promoter methylation of p16(INK4a), RARB2 and SEMA3B in bronchial aspirates from patients with suspected lung cancer. Int J Cancer. 2005;116:720–5.CrossRefGoogle Scholar
  19. 19.
    Kuroki T, Trapasso F, Yendamuri S, Matsuyama A, Alder H, Williams NN, et al. Allelic loss on chromosome 3p21.3 and promoter hypermethylation of semaphorin 3B in non-small cell lung cancer. Cancer Res. 2003;63:3352–5.Google Scholar
  20. 20.
    Ito M, Ito G, Kondo M, Uchiyama M, Fukui T, Mori S, et al. Frequent inactivation of RASSF1A, BLU, and SEMA3B on 3p21.3 by promoter hypermethylation and allele loss in non-small cell lung cancer. Cancer Lett. 2005;225:131–9.CrossRefGoogle Scholar
  21. 21.
    Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116:281–97.CrossRefGoogle Scholar
  22. 22.
    Gao X, Qiao Y, Han D, Zhang Y, Ma N. Enemy or partner: relationship between intronic micrornas and their host genes. IUBMB Life. 2012;64:835–40.CrossRefGoogle Scholar
  23. 23.
    Taft RJ, Pang KC, Mercer TR, Dinger M, Mattick JS. Non-coding RNAs: regulators of disease. J Pathol. 2010;220:126–39.CrossRefGoogle Scholar
  24. 24.
    Iyer MK, Niknafs YS, Malik R, Singhal U, Sahu A, Hosono Y, et al. The landscape of long noncoding RNAs in the human transcriptome. Nat Genet. 2015;47:199–208.CrossRefGoogle Scholar
  25. 25.
    Schmitt AM, Chang HY. Long noncoding RNAs in cancer pathways. Cancer Cell. 2016;29:452–63.CrossRefGoogle Scholar
  26. 26.
    Amin V, Harris RA, Onuchic V, Jackson AR, Charnecki T, Paithankar S, et al. Epigenomic footprints across 111 reference epigenomes reveal tissue-specific epigenetic regulation of lincRNAs. Nat Commun. 2015;6:6370.CrossRefGoogle Scholar
  27. 27.
    Wang Z, Yang B, Zhang M, Guo W, Wu Z, Wang Y, et al. LncRNA epigenetic landscape analysis identifies EPIC1 as an oncogenic lncRNA that interacts with MYC and promotes cell-cycle progression in cancer. Cancer Cell. 2018;33:706–20.CrossRefGoogle Scholar
  28. 28.
    Siewert JR, Stein HJ. Classification of adenocarcinoma of the oesophagogastric junction. Br J Surg. 1998;85:1457–9.CrossRefGoogle Scholar
  29. 29.
    Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 2001;25:402–8.CrossRefGoogle Scholar
  30. 30.
    Yu L, Liu C, Vandeusen J, Becknell B, Dai Z, Wu YZ, et al. Global assessment of promoter methylation in a mouse model of cancer identifies ID4 as a putative tumor-suppressor gene in human leukemia. Nat Genet. 2005;37:265–74.CrossRefGoogle Scholar
  31. 31.
    Loginov VI, Khodyrev DS, Pronina IV, Maliukova AV, Kazubskaia TP, Ermilova VD, et al. Two CpG-islands of SEMA3B gene: methylation in clear cell renal cell carcinoma. Mol Biol (Mosk). 2009;43:1088–92.Google Scholar
  32. 32.
    Dhar SS, Zhao D, Lin T, Gu B, Pal K, Wu SJ, et al. MLL4 Is Required to Maintain Broad H3K4me3 Peaks and Super-Enhancers at Tumor Suppressor Genes. Mol Cell. 2018;70:825–41.CrossRefGoogle Scholar
  33. 33.
    Ananthanarayanan M, Li Y, Surapureddi S, Balasubramaniyan N, Ahn J, Goldstein JA, et al. Histone H3K4 trimethylation by MLL3 as part of ASCOM complex is critical for NR activation of bile acid transporter genes and is downregulated in cholestasis. Am J Physiol Gastrointest Liver Physiol. 2011;300:G771–81.CrossRefGoogle Scholar
  34. 34.
    Neufeld G, Kessler O. The semaphorins: versatile regulators of tumour progression and tumour angiogenesis. Nat Rev Cancer. 2008;8:632–45.CrossRefGoogle Scholar
  35. 35.
    Tse C, Xiang RH, Bracht T, Naylor SL. Human Semaphorin 3B (SEMA3B) located at chromosome 3p21.3 suppresses tumor formation in an adenocarcinoma cell line. Cancer Res. 2002;62:542–6.Google Scholar
  36. 36.
    Griffiths-Jones S, Grocock RJ, van Dongen S, Bateman A, Enright AJ. miRBase: microRNA sequences, targets and gene nomenclature. Nucleic Acids Res. 2006;34:D140–4.CrossRefGoogle Scholar
  37. 37.
    Wang G, Wang Y, Shen C, Huang YW, Huang K, Huang TH, et al. RNA polymerase II binding patterns reveal genomic regions involved in microRNA gene regulation. PLoS One. 2010;5:e13798.CrossRefGoogle Scholar
  38. 38.
    Baskerville S, Bartel DP. Microarray profiling of microRNAs reveals frequent coexpression with neighboring miRNAs and host genes. RNA. 2005;11:241–7.CrossRefGoogle Scholar
  39. 39.
    Bach DH, Lee SK. Long noncoding RNAs in cancer cells. Cancer Lett. 2018;419:152–66.CrossRefGoogle Scholar
  40. 40.
    Wei S, Wang L, Zhang L, Li B, Li Z, Zhang Q, et al. ZNF143 enhances metastasis of gastric cancer by promoting the process of EMT through PI3K/AKT signaling pathway. Tumour Biol. 2016;37:12813–21.CrossRefGoogle Scholar

Copyright information

© The International Gastric Cancer Association and The Japanese Gastric Cancer Association 2019

Authors and Affiliations

  • Wei Guo
    • 1
  • Xiaoliang Liang
    • 1
  • Lei Liu
    • 2
  • Yanli Guo
    • 1
  • Supeng Shen
    • 1
  • Jia Liang
    • 1
  • Zhiming Dong
    • 1
    Email author
  1. 1.Laboratory of Pathology, Hebei Cancer InstituteThe Fourth Hospital of Hebei Medical UniversityShijiazhuangChina
  2. 2.Department of Thoracic SurgeryThe Fourth Hospital of Hebei Medical UniversityShijiazhuangChina

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