Effects of RNA silencing of matrix metalloproteinase-2 on the growth of esophageal carcinoma cells in vivo

Shen,Yu‑Guang; Feng,Wen; Xu,Yi‑Jun; Jiao,Na‑Na; Sun,Da‑Qiang; Qu,Wen‑Dong; Tang,Quan; Xiong,Wei; Tang,Yang; Xia,Yu; Cai,Qing‑Yong; Liu,Da‑Xing; Zhang,Xun; Xu,Gang; Liang,Gui‑You

doi:10.3892/ol.2016.5542

March-2017 Volume 13 Issue 3

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March-2017 Volume 13 Issue 3

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Article Open Access

Effects of RNA silencing of matrix metalloproteinase-2 on the growth of esophageal carcinoma cells in vivo

Authors:
- Yu‑Guang Shen
- Wen Feng
- Yi‑Jun Xu
- Na‑Na Jiao
- Da‑Qiang Sun
- Wen‑Dong Qu
- Quan Tang
- Wei Xiong
- Yang Tang
- Yu Xia
- Qing‑Yong Cai
- Da‑Xing Liu
- Xun Zhang
- Gang Xu
- Gui‑You Liang
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Affiliations: Department of Thoracic and Cardiovascular Surgery, The First People’s Hospital of Zunyi, Zunyi, Guizhou 563003, P.R. China, Department of Pathology, Henan Tumor Hospital of Zhengzhou University, Zhengzhou, Henan 450000, P.R. China, Thoracic Department, Tianjin Chest Hospital, Tianjin 300051, P.R. China, Department of Thoracic and Cardiovascular Surgery, Affiliated Hospital of Zunyi Medical College, Guizhou 563003, P.R. China

Copyright: © Shen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Pages: 1119-1124
|
Published online on: December 27, 2016

https://doi.org/10.3892/ol.2016.5542
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Abstract

Esophageal carcinoma is one of the most common malignancies in China. Previous studies reported that matrix metalloproteinases (MMPs) have important roles in the progression and invasion of numerous types of solid tumors. Among the MMPs, MMP‑2 has been closely associated with tumor growth and invasion. In the present study, a short hairpin RNA (shRNA) lentiviral expression vector targeting the MMP‑2 gene was constructed in order to observe the inhibitory effect of MMP‑2 gene silencing on the growth of the KYSE150 esophageal carcinoma cell line in vivo. Three small hairpin RNA sequences targeting MMP‑2 were designed and cloned into lentiviral vectors. Following transfection of the lentiviral vectors into KTSE150 cells, MMP‑2 mRNA and protein expression levels were examined by reverse transcription‑quantitative polymerase chain reaction and western blotting, and the growth rate of cells was analyzed by MTT assays. Subsequently, tumor growth was assessed in nude mice. Lentivirus‑mediated RNA interference effectively inhibited the expression of MMP‑2 mRNA and protein in KYSE150 esophageal carcinoma cells, and suppressed the growth of esophageal carcinoma cells in vivo. The results of the present study suggested that lentivirus‑mediated gene therapy targeting MMP‑2 may be an attractive strategy for the treatment of esophageal carcinoma and justifies the performance of further studies on the application of lentivirus vectors to cancer gene therapy.

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1	Mukherjee S, Roth MJ, Dawsey SM, Yan W, RodriguezCanales J, Erickson HS, Hu N, Goldstein AM, Taylor PR, Richardson AM, et al: Increased matrix metalloproteinase activation in esophageal squamous cell carcinoma. J Transl Med. 8:912010. View Article : Google Scholar : PubMed/NCBI
2	Li Y, Ma J, Guo Q, Duan F, Tang F, Zheng P, Zhao Z and Lu G: Overexpression of MMP-2 and MMP-9 in esophageal squamous cell carcinoma. Dis Esophagus. 22:664–667. 2009. View Article : Google Scholar : PubMed/NCBI
3	Poincloux R, Lizárraga F and Chavrier P: Matrix invasion by tumour cells: A focus on MT1-MMP trafficking to invadopodia. J Cell Sci. 122:3015–3024. 2009. View Article : Google Scholar : PubMed/NCBI
4	Yadav L, Puri N, Rastogi V, Satpute P, Ahmad R and Kaur G: Matrix metalloproteinases and cancer-roles in threat and therapy. Asian Pac J Cancer Prev. 15:1085–1091. 2014. View Article : Google Scholar : PubMed/NCBI
5	Price SJ, Greaves DR and Watkins H: Identification of novel, functional genetic variants in the human matrix metalloproteinase-2 gene: Role of Sp1 in allele-specific transcriptional regulation. J Biol Chem. 276:7549–7558. 2001. View Article : Google Scholar : PubMed/NCBI
6	Vihinen P and Kähäri VM: Matrix metalloproteinases in cancer: Prognostic markers and therapeutic targets. Int J Cancer. 99:157–166. 2002. View Article : Google Scholar : PubMed/NCBI
7	Ellenrieder V, Alber B, Lacher U, Hendler SF, Menke A, Boeck W, Wagner M, Wilda M, Friess H, Büchler M, et al: Role of MT-MMPs and MMP-2 in pancreatic cancer progression. Int J Cancer. 85:14–20. 2000. View Article : Google Scholar : PubMed/NCBI
8	Wang Y, Hu C, Dong R, Huang X and Qiu H: Platelet-derived growth factor-D promotes ovarian cancer invasion by regulating matrix metalloproteinases 2 and 9. Asian Pac J Cancer Prev. 12:3367–3370. 2011.PubMed/NCBI
9	Li GH, Cui YS, Wu QY, Zhang XJ and Gao YF: Clinicopathologic significance of β-catenin and matrix metalloproteinase-2 expression in non-small cell lung cancer. Med Oncol. 30:4372013. View Article : Google Scholar : PubMed/NCBI
10	Izquierdo M: Short interfering RNAs as a tool for cancer gene therapy. Cancer Gene Ther. 12:217–227. 2005. View Article : Google Scholar : PubMed/NCBI
11	Pushparaj PN, Aarthi JJ, Manikandan J and Kumar SD: SiRNA, miRNA and shRNA: In vivo applications. J Dent Res. 87:992–1003. 2008. View Article : Google Scholar : PubMed/NCBI
12	Amarzguioui M, Rossi JJ and Kim D: Approaches for chemically synthesized siRNA and vector-mediated RNAi. FEBS Lett. 579:5974–5981. 2005. View Article : Google Scholar : PubMed/NCBI
13	Li CX, Parker A, Menocal E, Xiang S, Borodyansky L and Fruehauf JH: Delivery of RNA interference. Cell Cycle. 5:2103–2109. 2006. View Article : Google Scholar : PubMed/NCBI
14	Shen YG, Xu YJ, Shi ZL, Han HL, Sun DQ and Zhang X: Effects of RNAi-mediated matrix metalloproteinase-2 gene silencing on the invasiveness and adhesion of esophageal carcinoma cells, KYSE150. Dig Dis Sci. 57:32–37. 2012. View Article : Google Scholar : PubMed/NCBI
15	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. View Article : Google Scholar : PubMed/NCBI
16	Chen W, He Y, Zheng R, Zhang S, Zeng H, Zou X and He J: Esophageal cancer incidence and mortality in China, 2009. J Thorac Dis. 5:19–26. 2013.PubMed/NCBI
17	Pennathur A, Gibson MK, Jobe BA and Luketich JD: Oesophageal carcinoma. Lancet. 381:400–412. 2013. View Article : Google Scholar : PubMed/NCBI
18	Roy R, Yang J and Moses MA: Matrix metalloproteinases as novel biomarkers and potential therapeutic targets in human cancer. J Clin Oncol. 27:5287–5297. 2009. View Article : Google Scholar : PubMed/NCBI
19	Gong J, Zhu S, Zhang Y and Wang J: Interplay of VEGFa and MMP2 regulates invasion of glioblastoma. Tumour Biol. 34:11879–11885. 2014. View Article : Google Scholar
20	Honkavuori-Toivola M, Santala M, Soini Y, Turpeenniemi- Hujanen T and Talvensaari-Mattila A: Combination of strong MMP-2 and weak TIMP-2 immunostainings is a significant prognostic factor in endometrial carcinoma. Dis Markers. 35:261–266. 2013. View Article : Google Scholar : PubMed/NCBI
21	Seo JM, Park S and Kim JH: Leukotriene B4 receptor-2 promotes invasiveness and metastasis of ovarian cancer cells through signal transducer and activator of transcription 3 (STAT3)-dependent up-regulation of matrix metalloproteinase 2. J Biol Chem. 287:13840–13849. 2012. View Article : Google Scholar : PubMed/NCBI
22	Sun Y, Liu M, Yang B, Li B and Lu J: Role of siRNA silencing of MMP-2 gene on invasion and growth of laryngeal squamous cell carcinoma. Eur Arch Otorhinolaryngol. 265:1385–1391. 2008. View Article : Google Scholar : PubMed/NCBI
23	Maddirela DR, Kesanakurti D, Gujrati M and Rao JS: MMP-2 suppression abrogates irradiation-induced microtubule formation in endothelial cells by inhibiting αvβ3-mediated SDF-1/CXCR4 signaling. Int J Oncol. 42:1279–1288. 2013.PubMed/NCBI
24	Zamore PD: RNA interference: Listening to the sound of silence. Nat Struct Biol. 8:746–750. 2001. View Article : Google Scholar : PubMed/NCBI
25	Leng Q, Woodle MC, Lu PY and Mixson AJ: Advances in systemic siRNA delivery. Drugs Future. 34:7212009. View Article : Google Scholar : PubMed/NCBI
26	Lin CC, Hsu JT, Huang KL, Tang HK and Lai YK: Stable RNA interference in Spodoptera frugiperda cells by a DNA vector-based method. Biotechnol Lett. 28:271–277. 2006. View Article : Google Scholar : PubMed/NCBI
27	Manjunath N, Wu H, Subramanya S and Shankar P: Lentiviral delivery of short hairpin RNAs. Adv Drug Deliv Rev. 61:732–745. 2009. View Article : Google Scholar : PubMed/NCBI
28	Dykxhoorn DM, Novina CD and Sharp PA: Killing the messenger: Short RNAs that silence gene expression. Nat Rev Mol Cell Biol. 4:457–467. 2003. View Article : Google Scholar : PubMed/NCBI
29	Lin CC, Hsu JT, Huang KL, Tang HK and Lai YK: Stable RNA interference in Spodoptera frugiperda cells by a DNA vector-based method. Biotechnol Lett. 28:271–277. 2006. View Article : Google Scholar : PubMed/NCBI
30	Ginn SL, Alexander IE, Edelstein ML, Abedi MR and Wixon J: Gene therapy clinical trials worldwide to 2012 - an update. J Gene Med. 15:65–77. 2013. View Article : Google Scholar : PubMed/NCBI
31	Chira S, Jackson CS, Oprea I, Ozturk F, Pepper MS, Diaconu I, Braicu C, Raduly LZ, Calin GA and Berindan-Neagoe I: Progresses towards safe and efficient gene therapy vectors. Oncotarget. 6:30675–30703. 2015.PubMed/NCBI