Simultaneous knockdown of APRIL via multiple shRNAs reduces the malignancy of SW480 cells

Li,Haiquan; Shao,Keke; Wang,Jingchun; Wang,Guihua; Xu,Jin; Cao,Jinde; Ju,Shaoqing; Wang,Huimin

doi:10.3892/or.2012.1980

Simultaneous knockdown of APRIL via multiple shRNAs reduces the malignancy of SW480 cells

Authors:
- Haiquan Li
- Keke Shao
- Jingchun Wang
- Guihua Wang
- Jin Xu
- Jinde Cao
- Shaoqing Ju
- Huimin Wang
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Affiliations: Department of Clinical Laboratory Center, Affiliated Hospital of Nantong University, Nantong Medical College, Nantong University, Nantong 226001, P.R. China, Clinical Laboratory, Yancheng City Nο. 1 People's Hospital, Yancheng 224001, P.R. China
Published online on: August 22, 2012 https://doi.org/10.3892/or.2012.1980
Pages: 1613-1618

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Abstract

A proliferation-inducing ligand (APRIL) is a key factor involved in the tumor development and progression in some tumor tissues and cells. Its overexpression and as gene target in SW480 colon carcinoma cells was confirmed in our previous study. To seek a more potent way to treat colon carcinoma using a gene therapy method, herein, we constructed a multiple short hairpin RNA (shRNA) expression vector containing four shRNAs against the APRIL gene in SW480 cells. APRIL expression levels and cell biological behavior were detected after transfection with different kinds of vectors. As expected, we found that our multiple shRNA vector produced a more significant knockdown effect of APRIL than the vectors containing only one APRIL shRNA. Furthermore, our findings indicate that silencing APRIL expression in SW480 cells decreased their malignancy by reducing proliferation, invasion and adhesion, as well as inducing apoptosis. Based on our findings, vectors containing multiple shRNAs to silence the expression of APRIL may be exploited as a novel therapeutic strategy for tumors.

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1	Hahne M, Kataoka T, Schröter M, Hofmann K, Irmler M, Bodmer JL, Schneider P, Bornand T, Holler N, French LE, et al: APRIL, a new ligand of the tumor necrosis factor family, stimulates tumor cell growth. J Exp Med. 188:1185–1190. 1998. View Article : Google Scholar : PubMed/NCBI
2	Kimberley FC, van Bostelen L, Cameron K, Hardenberg G, Marquart JA, Hahne M and Medema JP: The proteoglycan (heparan sulfate proteoglycan) binding domain of APRIL serves as a platform for ligand multimerization and cross-linking. FASEB J. 23:1584–1595. 2009. View Article : Google Scholar : PubMed/NCBI
3	Deshayes F, Laprée G, Portier A, Richard Y, Pencalet P, Mahieu-Caputo D, Horellou P and Tsapis A: Abnormal production of the TNF-homologue APRIL increases the proliferation of human malignant glioblastoma cell lines via a specific receptor. Oncogene. 23:3005–3012. 2004. View Article : Google Scholar : PubMed/NCBI
4	Sun B, Wang H, Wang X, Huang H, Ding W, Jing R, Shi G and Zhu L: A proliferation-inducing ligand: a new biomarker for non-small cell lung cancer. Exp Lung Res. 35:486–500. 2009. View Article : Google Scholar : PubMed/NCBI
5	Mhawech-Fauceglia P, Kaya G, Sauter G, McKee T, Donze O, Schwaller J and Huard B: The source of APRIL up-regulation in human solid tumor lesions. J Leukoc Biol. 80:697–704. 2006.PubMed/NCBI
6	Planelles L, Medema JP, Hahne M and Hardenberg G: The expanding role of APRIL in cancer and immunity. Curr Mol Med. 8:829–844. 2008. View Article : Google Scholar : PubMed/NCBI
7	Wang F, Chen L, Mao ZB, Shao JG, Tan C and Huang WD: Lentivirus-mediated short hairpin RNA targeting the APRIL gene suppresses the growth of pancreatic cancer cells in vitro and in vivo. Oncol Rep. 20:135–139. 2008.PubMed/NCBI
8	Moreaux J, Veyrune JL, De Vos J and Klein B: APRIL is overexpressed in cancer: link with tumor progression. BMC Cancer. 9:832009. View Article : Google Scholar : PubMed/NCBI
9	Dillon SR, Gross JA, Ansell SM and Novak AJ: An APRIL to remember: novel TNF ligands as therapeutic targets. Nat Rev Drug Discov. 5:235–246. 2006. View Article : Google Scholar : PubMed/NCBI
10	Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE and Mello CC: Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature. 391:806–811. 1998. View Article : Google Scholar : PubMed/NCBI
11	Cogoni C and Macino G: Post-transcriptional gene silencing across kingdoms. Curr Opin Genet Dev. 10:638–643. 2000. View Article : Google Scholar : PubMed/NCBI
12	Sioud M: Therapeutic siRNAs. Trends Pharmacol Sci. 25:22–28. 2004. View Article : Google Scholar
13	Wang S, Shi Z, Liu W, Jules J and Feng X: Development and validation of vectors containing multiple siRNA expression cassettes for maximizing the efficiency of gene silencing. BMC Biotechnol. 6:502006. View Article : Google Scholar : PubMed/NCBI
14	Ding W, Wang J, Sun B, Ju S, Yuan H, Wang X, Wang Y and Wang H: APRIL knockdown suppresses migration and invasion of human colon carcinoma cells. Clin Biochem. 42:1694–1698. 2009. View Article : Google Scholar : PubMed/NCBI
15	Hendriks J, Planelles L, de Jong-Odding J, Hardenberg G, Pals ST, Hahne M, Spaargaren M and Medema JP: Heparan sulfate proteoglycan binding promotes APRIL-induced tumor cell proliferation. Cell Death Differ. 12:637–648. 2005. View Article : Google Scholar : PubMed/NCBI
16	Rennert P, Schneider P, Cachero TG, Thompson J, Trabach L, Hertig S, Holler N, Qian F, Mullen C, Strauch K, Browning JL, Ambrose C and Tschopp J: A soluble form of B cell maturation antigen, a receptor for the tumor necrosis factor family member APRIL, inhibits tumor cell growth. J Exp Med. 192:1677–1684. 2000. View Article : Google Scholar : PubMed/NCBI
17	Marsters SA, Yan M, Pitti RM, Haas PE, Dixit VM and Ashkenazi A: Interaction of the TNF homologues BLyS and APRIL with the TNF receptor homologues BCMA and TACI. Curr Biol. 10:785–788. 2000. View Article : Google Scholar : PubMed/NCBI
18	Wu Y, Bressette D, Carrell JA, Kaufman T, Feng P, Taylor K, Gan Y, Cho YH, Garcia AD, Gollatz E, et al: Tumor necrosis factor (TNF) receptor superfamily member TACI is a high affinity receptor for TNF family members APRIL and BLyS. J Biol Chem. 275:35478–35485. 2000. View Article : Google Scholar : PubMed/NCBI
19	Esko JD and Lindahl U: Molecular diversity of heparan sulfate. J Clin Invest. 108:169–173. 2001. View Article : Google Scholar : PubMed/NCBI
20	Gallagher JT: Heparan sulfate: growth control with a restricted sequence menu. J Clin Invest. 108:357–361. 2001. View Article : Google Scholar : PubMed/NCBI
21	Perrimon N and Bernfield M: Specificities of heparan sulphate proteoglycans in developmental processes. Nature. 404:725–728. 2000. View Article : Google Scholar : PubMed/NCBI
22	Hayashida K, Johnston DR, Goldberger O and Park PW: Syndecan-1 expression in epithelial cells is induced by transforming growth factor beta through a PKA-dependent pathway. J Biol Chem. 281:24365–24374. 2006. View Article : Google Scholar : PubMed/NCBI
23	Ding W, Ju S, Jiang S, Zhu L, Wang Y and Wang H: Reduced APRIL expression induces cellular senescence via a HSPG-dependent pathway. Pathol Oncol Res. 15:693–701. 2009. View Article : Google Scholar : PubMed/NCBI
24	Hannon GJ: RNA interference. Nature. 418:244–251. 2002. View Article : Google Scholar : PubMed/NCBI
25	Dykxhoorn DM: RNA interference as an anticancer therapy: a patent perspective. Expert Opin Ther Pat. 19:475–491. 2009. View Article : Google Scholar : PubMed/NCBI
26	Wang SL, Yao HH and Qin ZH: Strategies for short hairpin RNA delivery in cancer gene therapy. Expert Opin Biol Ther. 9:1357–1368. 2009. View Article : Google Scholar : PubMed/NCBI
27	Gou D, Weng T, Wang Y, Wang Z, Zhang H, Gao L, Chen Z, Wang P and Liu L: A novel approach for the construction of multiple shRNA expression vectors. J Gene Med. 9:751–763. 2007. View Article : Google Scholar : PubMed/NCBI
28	Yan Y, Zhang J, Guo JL, Huang W and Yang YZ: Multiple shRNA-mediated knockdown of TACE reduces the malignancy of HeLa cells. Cell Biol Int. 33:158–164. 2009. View Article : Google Scholar : PubMed/NCBI
29	Jemal A, Siegel R, Xu J and Ward E: Cancer statistics, 2010. CA Cancer J Clin. 60:277–300. 2010. View Article : Google Scholar
30	Center MM, Jemal A, Smith RA and Ward E: Worldwide variations in colorectal cancer. CA Cancer J Clin. 59:366–378. 2009. View Article : Google Scholar : PubMed/NCBI
31	Ortega J, Vigil CE and Chodkiewicz C: Current progress in targeted therapy for colorectal cancer. Cancer Control. 17:7–15. 2010.PubMed/NCBI
32	Markowitz SD and Bertagnolli MM: Molecular origins of cancer: Molecular basis of colorectal cancer. N Engl J Med. 361:2449–2460. 2009. View Article : Google Scholar : PubMed/NCBI
33	Jazag A, Kanai F, Ijichi H, Tateishi K, Ikenoue T, Tanaka Y, Ohta M, Imamura J, Guleng B, Asaoka Y, et al: Single small-interfering RNA expression vector for silencing multiple transforming growth factor-beta pathway components. Nucleic Acids Res. 33:e1312005. View Article : Google Scholar : PubMed/NCBI
34	ter Brake O, Konstantinova P, Ceylan M and Berkhout B: Silencing of HIV-1 with RNA interference: a multiple shRNA approach. Mol Ther. 14:883–892. 2006.PubMed/NCBI
35	Song J, Giang A, Lu Y, Pang S and Chiu R: Multiple shRNA expressing vector enhances efficiency of gene silencing. BMB Rep. 41:358–362. 2008. View Article : Google Scholar : PubMed/NCBI