Tumor Biology

, Volume 34, Issue 5, pp 2951–2959 | Cite as

Effects of CXCR4 gene silencing by lentivirus shRNA on proliferation of the EC9706 human esophageal carcinoma cell line

  • Dao-feng Wang
  • Ning Lou
  • Miao-zhen Qiu
  • Yong-bin Lin
  • Ying Liang
Research Article


CXCL12/CXCR4 has been studied as an important biomarker for many human malignancies, but studies are limited for esophageal squamous cell carcinoma (ESCC). In this study, an effective RNAi sequence targeting the CXCR4 gene was selected, a lentiviral shRNA vector was constructed to specifically silence CXCR4 expression in the EC9706 ESCC cell line, and the effects of CXCR4 silencing on cell growth in vitro and tumour growth in nude mice were then evaluated. The expression of CXCR4 in EC9706 was significantly downregulated after transfection with a lentiviral shRNA vector. The expression of the apoptosis-related gene Bcl-2 was decreased. In addition, after CXCR4 inhibition, cell growth was considerably inhibited, increased apoptosis in the EC9706 cells was found, the G0/G1 percentage was significantly increased, and the number of cells in S phase was reduced. Moreover, tumour growth in nude mice was inhibited. In conclusion, the downregulation of CXCR4 expression by transfection with a lentiviral shRNA vector in ESCC cells could inhibit tumour proliferation. Our data may provide an avenue for finding new ESCC treatments.


CXCL12 CXCR4 Esophageal squamous cell carcinoma RNA interference Proliferation 



We gratefully thank the staff members in the Department of Medical Oncology and Thoracic Surgery Oncology at Sun Yat-sen University Cancer Center for their suggestion and assistance.

Conflicts of interest



This work was supported by a grant from the Natural Science Foundation of the Guangdong Province of China (S2011040004502).


  1. 1.
    Lu SH. Alterations of oncogenes and tumor suppressor genes in esophageal cancer in China. Mutat Res. 2000;462:343–53.CrossRefPubMedGoogle Scholar
  2. 2.
    Aiuti A et al. Expression of CXCR4, the receptor for stromal cell-derived factor-1 on fetal and adult human lympho-hematopoietic progenitors. Eur J Immunol. 1999;29:1823–31.CrossRefPubMedGoogle Scholar
  3. 3.
    Jinquan T et al. CXC chemokine receptor 4 expression and stromal cell-derived factor-1 alpha-induced chemotaxis in CD4+ T lymphocytes are regulated by interleukin-4 and interleukin-10. Immunology. 2000;99:402–10.PubMedCentralCrossRefPubMedGoogle Scholar
  4. 4.
    Oonakahara K, Matsuyama W, Higashimoto I, Kawabata M, Arimura K, Osame M. Stromal-derived factor-1alpha/CXCL12-CXCR 4 axis is involved in the dissemination of NSCLC cells into pleural space. Am J Respir Cell Mol Biol. 2004;30:671–7.CrossRefPubMedGoogle Scholar
  5. 5.
    Yasumoto K et al. Role of the CXCL12/CXCR4 axis in peritoneal carcinomatosis of gastric cancer. Cancer Res. 2006;66:2181–7.CrossRefPubMedGoogle Scholar
  6. 6.
    Sasaki K et al. Expression of CXCL12 and its receptor CXCR4 in esophageal squamous cell carcinoma. Oncol Rep. 2009;21:65–71.PubMedGoogle Scholar
  7. 7.
    Ganju RK et al. The alpha-chemokine, stromal cell-derived factor-1alpha, binds to the transmembrane G-protein-coupled CXCR-4 receptor and activates multiple signal transduction pathways. J Bio Chem. 1998;273:23169–75.CrossRefGoogle Scholar
  8. 8.
    Balkwill F. Cancer and the chemokine network. Nat Rev Cancer. 2004;4:540–50.CrossRefPubMedGoogle Scholar
  9. 9.
    Juarez J, Bradstock KF, Gottlieb DJ, Bendall LJ. Effects of inhibitors of the chemokine receptor CXCR4 on acute lymphoblastic leukemia cells in vitro. Leukemia. 2003;17:1294–300.CrossRefPubMedGoogle Scholar
  10. 10.
    Liu G et al. Critical role of SDF-1α-induced progenitor cell recruitment and macrophage VEGF production in the experimental corneal neovascularization. Mol Vis. 2011;17:2129–38.PubMedCentralPubMedGoogle Scholar
  11. 11.
    Yoon O, Roh J. Downregulation of KLF4 and the Bcl-2/Bax ratio in advanced epithelial ovarian cancer. Oncol Lett. 2012;4:1033–6.PubMedCentralPubMedGoogle Scholar
  12. 12.
    De Cesare M et al. Enhanced antitumour efficacy of gimatecan in combination with Bcl-2 antisense oligonucleotide in human melanoma xenografts. Eur J Cancer. 2005;41:1213–22.CrossRefPubMedGoogle Scholar
  13. 13.
    Peled A et al. The chemokine SDF-1 activates the integrins LFA-1, VLA-4, and VLA-5 on immature human CD34(+) cells: role in transendothelial/stromal migration and engraftment of NOD/SCID mice. Blood. 2000;95:3289–96.PubMedGoogle Scholar
  14. 14.
    Lee Y et al. Enhancement of intracellular signaling associated with hematopoietic progenitor cell survival in response to SDF-1/CXCL12 in synergy with other cytokines. Blood. 2002;99:4307–17.CrossRefPubMedGoogle Scholar
  15. 15.
    Brand S et al. CXCR4 and CXCL12 are inversely expressed in colorectal cancer cells and modulate cancer cell migration, invasion and MMP-9 activation. Exp Cell Res. 2005;310:117–30.CrossRefPubMedGoogle Scholar
  16. 16.
    Müller A et al. Involvement of chemokine receptors in breast cancer metastasis. Nature. 2001;410(6824):50–6.CrossRefPubMedGoogle Scholar
  17. 17.
    Wagner PL et al. CXCL12 and CXCR4 in adenocarcinoma of the lung: association with metastasis and survival. J Thorac Cardiovasc Surg. 2009;137:615–21.CrossRefPubMedGoogle Scholar
  18. 18.
    Sehgal A, Keener C, Boynton AL, Warrick J, Murphy GP. CXCR4, a chemokine receptor, is overexpressed in and required for proliferation of glioblastoma tumor cells. J Surg Oncol. 1998;69:99–104.CrossRefPubMedGoogle Scholar
  19. 19.
    Sasaki K et al. Expression of CXCL12 and its receptor CXCR4 correlates with lymph node metastasis in submucosal esophageal cancer. J Surg Oncol. 2008;97:433–8.CrossRefPubMedGoogle Scholar
  20. 20.
    Kaifi JT et al. Tumor-cell homing to lymph nodes and bone marrow and CXCR4 expression in esophageal cancer. J Natl Cancer Inst. 2005;97:1840–7.CrossRefPubMedGoogle Scholar
  21. 21.
    Li M, Rossi JJ. Lentiviral vector delivery of siRNA and shRNA encoding genes into cultured and primary hematopoietic cells. Methods Mol Biol. 2005;309:261–72.PubMedGoogle Scholar
  22. 22.
    Lapteva N, Yang AG, Sanders DE, Strube RW, Chen SY. CXCR4 knockdown by small interfering RNA abrogates breast tumor growth in vivo. Cancer Gene Ther. 2005;12:84–9.CrossRefPubMedGoogle Scholar
  23. 23.
    Wang DF, Lou N, Zeng CG, Zhang X, Chen FJ. Expression of CXCL12/CXCR4 and its correlation to prognosis in esophageal squamous cell carcinoma. Ai Zheng. 2009;28:154–8.PubMedGoogle Scholar
  24. 24.
    Wei L, Kong PY, Chen XH, Peng XG, Zeng DF, Chang C, et al. Effects of anti-CXCR4 monoclonal antibody on adhesion and proliferation of human acute myelocytic leukemia cell line HL-60. Ai Zheng. 2004;23:1273–7.PubMedGoogle Scholar
  25. 25.
    Hernández-López C, Varas A, Sacedón R, Jiménez E, Muñoz JJ, Zapata AG, et al. Stromal cell-derived factor 1/CXCR4 signaling is critical for early human T-cell development. Blood. 2002;99:546–54.CrossRefPubMedGoogle Scholar
  26. 26.
    Kryczek I et al. The chemokine SDF-1/CXCL12 contributes to T lymphocyte recruitment in human pre-ovulatory follicles and coordinates with lymphocytes to increase granulosa cell survival and embryo quality. Am J Reprod Immunol. 2005;54:270–83.CrossRefPubMedGoogle Scholar
  27. 27.
    Agui T, McConkey DJ, Tanigawa N. Comparative study of various biological parameters, including expression of survivin, between primary and metastatic human colonic adenocarcinomas. Anticancer Res. 2002;22:1769–76.PubMedGoogle Scholar
  28. 28.
    Tsai ST, Jin YT, Leung HW, Wang ST, Tsao CJ, Su IJ. Bcl-2 and proliferating cell nuclear antigen (PCNA) expression correlates with subsequent local recurrence in nasopharyngeal carcinomas. Anticancer Res. 1998;18:2849–54.PubMedGoogle Scholar
  29. 29.
    Lu CL, Ji Y, Ge D, Guo J, Ding JY. The expression of CXCR4 and its relationship with matrix metalloproteinase-9/vascular endothelial growth factor in esophageal squamous cell cancer. Dis Esophagus. 2011;24:283–90.CrossRefPubMedGoogle Scholar
  30. 30.
    Gros SJ, Graeff H, Drenckhan A, Kurschat N, Blessmann M, Rawnaq T, et al. CXCR4/SDF-1α-mediated chemotaxis in an in vivo model of metastatic esophageal carcinoma. In Vivo. 2012;26:711–8.PubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2013

Authors and Affiliations

  • Dao-feng Wang
    • 1
    • 2
  • Ning Lou
    • 1
    • 2
  • Miao-zhen Qiu
    • 1
    • 3
  • Yong-bin Lin
    • 1
    • 4
  • Ying Liang
    • 1
    • 3
  1. 1.State Key Laboratory of Oncology in South ChinaGuangzhouPeople’s Republic of China
  2. 2.Department of ICUSun Yat-Sen University Cancer CenterGuangzhouPeople’s Republic of China
  3. 3.Department of Medical OncologySun Yat-Sen University Cancer CenterGuangzhouPeople’s Republic of China
  4. 4.Department of Thoracic SurgerySun Yat-Sen University Cancer CenterGuangzhouPeople’s Republic of China

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