RNAi targeting CXCR4 inhibits proliferation and invasion of esophageal carcinoma cells

Diagnostic Pathology

Xiaoyan Xuan²  Yuling Zheng¹  Hong Xu⁴  Ping Gao³  Linping Pian⁶  Yanfang Mi⁵  Tao Wang⁷ 

[1] Henan University of TCM, Zhengzhou, P.R. China;College of Basic Medical Sciences, Zhengzhou University, Zhengzhou, P.R. China;Department of Hemato-tumor, The First Affiliated Hospital of Henan College, University of TCM, Zhengzhou, P.R. China;Henan Tumor Institute, Zhengzhou, P.R. China;Department of Otolaryngology Head and Neck Surgery, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou P.R. China;Department of Ultrasound, The First Affiliated Hospital College, TCM of Henan, Zhengzhou P.R. China;The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, P.R. China

关键词: Invasion;    Proliferation;    CXCR4;    Esophageal carcinoma;   

Others  :  806507
DOI  :  10.1186/1746-1596-8-104

 received in 2013-05-23, accepted in 2013-06-03,  发布年份 2013

【 摘 要 】

CXC chemokine receptor 4 was found to be expressed by many different types of human cancers and its expression has been correlated with tumor aggressiveness, poor prognosis and resistance to chemotherapy. However the effect of CXCR4 on the esophageal carcinoma cells remains unclear, the present study explored the effects of CXCR4 siRNA on proliferation and invasion of esophageal carcinoma KYSE-150 and TE-13 cells. Two siRNA sequence targeting CXCR4 gene were constructed and then were transfected into KYSE-150 and TE-13 cells by Lipofectamine™2000. Changes of CXCR4 mRNA and protein were analyzed by qRT-PCR and Western blot. Effect of CXCR4 siRNA on KYSE-150 and TE-13 cells proliferation was determined by MTT. Transwell invasion assay was used to evaluate the invasion and metastasis of KYSE-150 and TE-13 cells. Tumor growth was assessed by subcutaneous inoculation of cells into BALB/c nude mice. qRT-PCR and Western blot demonstrate that the expression level of CXCR4 gene were obviously decreased in KYSE-150 and TE-13 cells transfected with CXCR4 targeting siRNA expression vectors. The average amount of cells transfected with CXCR4 siRNA penetrating Matrigel was significantly decreased (p<0.05). Injection of CXCR4 siRNA transfected cells inhibited tumor growth in a xenograft model compared with blank and negative control groups (p <0.05). CXCR4 silenced by siRNA could suppress the proliferation, invasion and metastasis of esophageal carcinoma cell lines KYSE-150 and TE-13 in vitro and in vivo. The results provide a theoretical and experimental basis for the gene therapy of ESCC using RNAi technology based on CXCR4 target site.

Virtual slides

The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/3502376691001138 webcite

【 授权许可】

   
2013 Wang et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

Figure 8.

【 参考文献 】

• [1]Siegel R, Naishadham D, Jemal A: Cancer statistics. CA Cancer J Clin 2012, 62(1):10-29.
• [2]Parkin DM, Pisani P, Ferlay J: Estimates of the worldwide incidence of 25 major cancers in 1990. Int J Cancer 1990, 80:827-841.
• [3]Montesano R, Hollstein M, Hainant P: Genetic alterations in esophageal cancer and their relevance to etiology and pathogenesis: a review. Int J Cancer 1996, 69:225-235.
• [4]Burger JA, Kipps TJ: CXCR4: a key receptor in the crosstalk between tumor cells and their microenvironment. Blood 2006, 107:1761-1767.
• [5]Murdoch C: CXCR4: chemokine receptor extraordinaire. Immunol Rev 2000, 177:175-184.
• [6]Duda DG, Kozin SV, Kirkpatrick ND, Xu L, Fukumura D, et al.: CXCL12 (SDF1-α) - CXCR4/CXCR7 pathway inhibition: an emerging sensitizer for anticancer therapies. Clin Cancer Res 2011, 17:2074-2080.
• [7]Muller A, Homey B, Soto H, Ge N, Catron D, et al.: Involvement of chemokine receptors in breast cancer metastasis. Nature 2001, 410:50-56.
• [8]Liang Z, Yoon Y, Votaw J, Goodman MM, Williams L, et al.: Silencing of CXCR4 blocks breast cancer metastasis. Cancer Res 2005, 65:967-971.
• [9]Berghuis D, Santos SJ, Baelde HJ, Taminiau AH, Egeler RM, et al.: Pro-inflammatory chemokine-chemokine receptor interactions within the Ewing sarcoma microenvironment determine CD8(+) T-lymphocyte infiltration and affect tumour progression. J Pathol 2011, 223:347-357.
• [10]Wang L, Wang Z, Yang B, Yang Q, Wang L, et al.: CXCR4 nuclear localization follows binding of its ligand SDF-1 and occurs in metastatic but not primary renal cell carcinoma. Oncol Rep 2009, 22:1333-1339.
• [11]Wang SC, Lin JK, Wang HS, Yang SH, Li AF, et al.: Nuclear expression of CXCR4 is associated with advanced colorectal cancer. Int J Colorectal Dis 2010, 25:1185-1191.
• [12]Namlos HM, Kresse SH, Muler CR, Henriksen J, Holdhus R, et al.: Global gene expression profiling of human osteosarcomas reveals metastasis-association chemokine pattern. Sarcoma 2012, 2012:639038.
• [13]Baumhoer D, Smida J, Zillmer S, Rosemann M, Atkinson MJ, et al.: Strong expression of CXCL12 is associated with a favorable outcome in osteosarcoma. Mod Pathol 2012, 25:522-528.
• [14]Grymula K, Tarnowski M, Wysoczynski M, Drukala J, Barr FG, et al.: Overlapping and distinct role of CXCR7-SDF-1/ITAC and CXCR4-SDF-1 axes in regulating metastatic behavior of human rhabdomyosarcomas. Int J Cancer 2010, 127:2554-2568.
• [15]Furusato B, Mohamed A, Uhlen M, Rhim JS: CXCR4 and cancer. Pathol Int 2010, 60:497-505.
• [16]Tanaka T, Bai Z, Srinoulprasert Y, Yang BG, Hayasaka H, et al.: Chemokines in tumor progression and metastasis. Cancer Sci 2005, 96:317-322.
• [17]Redjal N, Chan JA, Segal RA, Kung AL: CXCR4 inhibition synergizes with cytotoxic chemotherapy in gliomas. Clin Cancer Res 2006, 12:6765-6771.
• [18]Taichman RS, Cooper C, Keller ET, Pienta KJ, Taichman NS, et al.: Use of the stromal cell-derived factor-1/CXCR4 pathway in prostate cancer metastasis to bone. Cancer Res 2002, 62:1832-1837.
• [19]Azab AK, Runnels JM, Pitsillides C, Moreau AS, Azab F, et al.: CXCR4 inhibitor AMD3100 disrupts the interaction of multiple myeloma cells with the bone marrow microenvironment and enhances their sensitivity to therapy. Blood 2009, 113:4341-4351.
• [20]Kurtova AV, Tamayo AT, Ford RJ, Burger JA: Mantle cell lymphoma cells express high levels of CXCR4, CXCR5, and VLA-4 (CD49d): importance for interactions with the stromal microenvironment and specific targeting. Blood 2009, 113:4604-4613.
• [21]Li JK, Yu L, Shen Y, Zhou LS, Wang YC, et al.: Inhibition of CXCR4 activity with AMD3100 decreases invasion of human colorectal cancer cells in vitro. World J Gastroenterology 2008, 14:2308-2313.
• [22]Lee HJ, Kim SW, Kim HY, Li S, Yun HJ, et al.: Chemokine receptor CXCR4 expression, function, and clinical implications in gastric cancer. Int J Oncol 2009, 34(2):473-480.
• [23]Peled A, Wald O, Burger J: Development of novel CXCR4-based therapeutics. Expert Opin Investig Drugs 2012, 21:341-353.
• [24]Hannon GJ: A interference. Nature 2002, 418:244-251.
• [25]Tuschl T, Borkhardt A: Small interfering RNAs: a revolutionary tool for the analysis of gene function and gene therapy. Mol Interventions 2002, 2:158-167.
• [26]Semizarov D, Frost L, Sarthy A, Kroeger P, Halbert DN, et al.: Specificity of short interfering RNA determined through gene expression signatures. Proc Natl Acad Sci 2003, 11:6347-6352.
• [27]Jackson AL, Bartz SR, Schelter J, Kobayashi SV, Burchard J, et al.: Expression profiling revealsoff-target gene regulation by RNAi. Nat Biotechnol 2003, 21:635-637.
• [28]Ghildiyal M, Zamore PD: Small silencing RNAs: an expanding universe. Nat Rev Genet 2009, 10:94-108.
• [29]Kim VN, Han J, Siomi MC: Biogenesis of small RNAs in animals. Nat Rev Mol Cell Biol 2009, 10:126-139.
• [30]Czech B, Hannon GJ: Small RNA sorting: matchmaking for Argonautes. Nat Rev Genet 2001, 12:19-31.
• [31]Kastenmeier A, Gonzales H, Gould JC: Robotic applications in the treatment of diseases of the esophagus. Surg Laparosc Endosc Percutan Tech 2012, 22(4):304-309.
• [32]Palanivelu C, Prakash A, Senthilkumar R, Senthilnathan P, Parthasarathi R, et al.: minimally invasive esophagectomy: thoracoscopic mobilization of the esophagus and medi-astinal lymphadenectomy in prone position-experience of 130 patients. J Am Coll Surg 2006, 203(1):7-16.