Targeting antitumor effect of rhTNF-α fusion protein mediated by matrix metalloproteinase-2

Shao,Xin; Ren,Hui; Wang,Yue-Li; Wang,Fa; Hou,Gan; Huang,Di-Nan

doi:10.3892/or.2014.3616

Targeting antitumor effect of rhTNF-α fusion protein mediated by matrix metalloproteinase-2

Authors:
- Xin Shao
- Hui Ren
- Yue-Li Wang
- Fa Wang
- Gan Hou
- Di-Nan Huang
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Affiliations: Institute of Biochemistry and Molecular Biology, Guangdong Medical College, Zhanjiang, Guangdong 524023, P.R. China, Department of Clinical Biochemistry, Guangdong Medical College, Dongguan, Guangdong 523808, P.R. China
Published online on: November 24, 2014 https://doi.org/10.3892/or.2014.3616
Pages: 810-818

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Abstract

The aim of this study was to examine the tumor therapy, targeting effects and side effects of tumor‑targeting rhTNF-α fusion protein mediated by matrix metalloproteinase-2 in an animal model in order to provide experimental data for future development of drugs. The median lethal dose (LD50) was obtained from acute toxicity experiments. The A549 lung cancer xenograft model was established, and then randomly divided into the saline, standard substance, and low‑, middle‑ and high‑dose fusion protein experiment groups. Each group was administered drugs for 18 days. The length and width of the xenografts were measured every three days, after which the xenograft growth curve was drawn. The mice were sacrificed in each group following treatment and the tumor volume and weight were measured. The targeting, effectiveness and toxicity of the transformed fusion protein, and pathological changes of tumor and organ tissues were examined by hematoxylin and eosin (H&E) staining. Additionally, biochemical markers were used to detect damage of various organs after protein processing. Cell apoptosis and angiogenesis were determined using terminal deoxynucleotidyltransferase-mediated dUTP nick end‑labeling (TUNEL) testing and immunohistochemistry, respectively, in different dose groups. Tumor growth was markedly retarded in the high‑dose experimental and standard hTNF-α groups with antitumor rates of 85.91 and 72.25%, respectively, as compared with the control group. Furthermore, the tumor tissue showed obvious apoptosis (the apoptotic index was 78.78 and 66.65%, respectively) and pathological changes in the high‑dose experimental and standard hTNF-α groups. Tumor angiogenesis in each fusion protein group was inhibited (P<0.01) and the biochemical markers of various organs were greatly reduced in the high‑dose experimental group (P<0.05). This finding indicated that slight toxic effects of fusion proteins were evident for the heart, liver and kidney. The reforming fusion protein can therefore target tumor tissues and efficiently kill tumor cells, with few side effects.

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1	Jo SK, Hong JY, Park HJ and Lee SK: Anticancer activity of novel daphnane diterpenoids from Daphne genkwa through cell-cycle arrest and suppression of Akt/STAT/Src signalings in human lung cancer cells. Biomol Ther (Seoul). 20:513–519. 2012. View Article : Google Scholar
2	Wangari-Talbot J and Hopper-Borge E: Drug resistance mechanisms in non-small cell lung carcinoma. J Can Res Updates. 2:265–282. 2013.
3	Zhao P, Dai M, Chen W and Li N: Cancer trends in China. Jpn J Clin Oncol. 40:281–285. 2010. View Article : Google Scholar : PubMed/NCBI
4	Zhang Y, Wu JZ, Zhang JY, et al: Detection of circulating vascular endothelial growth factor and matrix metalloproteinase-9 in non-small cell lung cancer using Luminex multiplex technology. Oncol Lett. 7:499–506. 2014.PubMed/NCBI
5	Wang S: The promise of cancer therapeutics targeting the TNF-related apoptosis-inducing ligand and TRAIL receptor pathway. Oncogene. 27:6207–6215. 2008. View Article : Google Scholar : PubMed/NCBI
6	Carswell EA, Old LJ, Kassel RL, Green S, Fiore N and Williamson B: An endotoxin-induced serum factor that causes necrosis of tumors. Proc Natl Acad Sci USA. 72:3666–3670. 1975. View Article : Google Scholar : PubMed/NCBI
7	Old LJ: Tumor necrosis factor (TNF). Science. 230:630–632. 1985. View Article : Google Scholar : PubMed/NCBI
8	Van Roy M, Wielockx B, Baker A and Libert C: The use of tissue inhibitors of matrix metalloproteinases to increase the efficacy of a tumor necrosis factor/interferon gamma antitumor therapy. Cancer Gene Ther. 14:372–379. 2007. View Article : Google Scholar : PubMed/NCBI
9	Alexander HR Jr, Bartlett DL and Libutti SK: Current status of isolated hepatic perfusion with or without tumor necrosis factor for the treatment of unresectable cancers confined to liver. Oncologist. 5:416–424. 2000. View Article : Google Scholar : PubMed/NCBI
10	Grunhagen DJ, Brunstein F, ten Hagen TL, van Geel AN, de Wilt JH and Eggermont AM: TNF-based isolated limb perfusion: a decade of experience with antivascular therapy in the management of locally advanced extremity soft tissue sarcomas. Cancer Treat Res. 120:65–79. 2004. View Article : Google Scholar : PubMed/NCBI
11	Grunhagen DJ, de Wilt JH, ten Hagen TL and Eggermont AM: Technology insight: Utility of TNF-alpha-based isolated limb perfusion to avoid amputation of irresectable tumors of the extremities. Nat Clin Pract Oncol. 3:94–103. 2006. View Article : Google Scholar : PubMed/NCBI
12	Son KJ, Shin DS, Kwa T, Gao Y and Revzin A: Micropatterned sensing hydrogels integrated with reconfigurable microfluidics for detecting protease release from cells. Anal Chem. 85:11893–11901. 2013. View Article : Google Scholar : PubMed/NCBI
13	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
14	Weng Y, Cai M, Zhu J, et al: Matrix metalloproteinase activity in early-stage lung cancer. Onkologie. 36:256–259. 2013. View Article : Google Scholar : PubMed/NCBI
15	Kao SJ, Su JL, Chen CK, et al: Osthole inhibits the invasive ability of human lung adenocarcinoma cells via suppression of NF-κB-mediated matrix metalloproteinase-9 expression. Toxicol Appl Pharmacol. 261:105–115. 2012. View Article : Google Scholar : PubMed/NCBI
16	Leinonen T, Pirinen R, Bohm J, Johansson R, Ropponen K and Kosma VM: Expression of matrix metalloproteinases 7 and 9 in non-small cell lung cancer. Relation to clinicopathological factors, beta-catenin and prognosis. Lung Cancer. 51:313–321. 2006. View Article : Google Scholar : PubMed/NCBI
17	Siejka A, Barabutis N and Schally AV: GHRH antagonist inhibits focal adhesion kinase (FAK) and decreases expression of vascular endothelial growth factor (VEGF) in human lung cancer cells in vitro. Peptides. 37:63–68. 2012. View Article : Google Scholar : PubMed/NCBI
18	Shao W, Wang W, Xiong XG, et al: Prognostic impact of MMP-2 and MMP-9 expression in pathologic stage IA non-small cell lung cancer. J Surg Oncol. 104:841–846. 2011. View Article : Google Scholar : PubMed/NCBI
19	Qian Q, Wang Q, Zhan P, et al: The role of matrix metalloproteinase 2 on the survival of patients with non-small cell lung cancer: a systematic review with meta-analysis. Cancer Invest. 28:661–669. 2010. View Article : Google Scholar : PubMed/NCBI
20	Desmard M, Amara N, Lanone S, Motterlini R and Boczkowski J: Carbon monoxide reduces the expression and activity of matrix metalloproteinases 1 and 2 in alveolar epithelial cells. Cell Mol Biol (Noisy-le-grand). 51:403–408. 2005.
21	Chauhan V, Breznan D, Thomson E, Karthikeyan S and Vincent R: Effects of ambient air particles on the endothelin system in human pulmonary epithelial cells (A549). Cell Biol Toxicol. 21:191–205. 2005. View Article : Google Scholar : PubMed/NCBI
22	Pardo A, Gibson K, Cisneros J, et al: Up-regulation and profibrotic role of osteopontin in human idiopathic pulmonary fibrosis. PLoS Med. 2:e2512005. View Article : Google Scholar : PubMed/NCBI
23	Weidner N: Current pathologic methods for measuring intratumoral microvessel density within breast carcinoma and other solid tumors. Breast Cancer Res Treat. 36:169–180. 1995. View Article : Google Scholar : PubMed/NCBI
24	Mendolicchio GL and Ruggeri ZM: New perspectives on von Willebrand factor functions in hemostasis and thrombosis. Semin Hematol. 42:5–14. 2005. View Article : Google Scholar : PubMed/NCBI
25	Cao C, Qi Y, Chen W, Zhu Y and Chen X: Effects of IKKɛ on oxidised low-density lipoprotein-induced injury in vascular endothelial cells. Heart Lung Circ. 22:366–372. 2013. View Article : Google Scholar
26	Ulger H, Karabulut AK and Pratten MK: Labelling of rat endothelial cells with antibodies to vWF, RECA-1, PECAM-1, ICAM-1, OX-43 and ZO-1. Anat Histol Embryol. 31:31–35. 2002. View Article : Google Scholar : PubMed/NCBI
27	Ruggeri ZM: Structure of von Willebrand factor and its function in platelet adhesion and thrombus formation. Best Pract Res Clin Haematol. 14:257–279. 2001. View Article : Google Scholar : PubMed/NCBI
28	Cai W, Kerner ZJ, Hong H and Sun J: Targeted cancer therapy with tumor necrosis factor-alpha. Biochem Insights. 2008:15–21. 2008.PubMed/NCBI
29	Gaughan EM and Costa DB: Genotype-driven therapies for non-small cell lung cancer: focus on EGFR, KRAS and ALK gene abnormalities. Ther Adv Med Oncol. 3:113–125. 2011. View Article : Google Scholar : PubMed/NCBI
30	Cooke SP, Pedley RB, Boden R, Begent RH and Chester KA: In vivo tumor delivery of a recombinant single chain Fv::tumor necrosis factor-alpha fusion [correction of factor: a fusion] protein. Bioconjug Chem. 13:7–15. 2002. View Article : Google Scholar : PubMed/NCBI