Article
Antitumor effect and mechanism of action of a tumor-targeting recombinant human tumor necrosis factor-α fusion protein mediated by urokinase
- Authors:
- You‑Chao Dai
- Si‑Min Yang
- Xin Wang
- Yong‑Jun Zhou
- Gan Hou
- Di‑Nan Huang
-
View Affiliations / Copyright
Affiliations:
Institute of Medical Inspection, Guangdong Medical College, Zhanjiang, Guangdong 523808, P.R. China, Department of Clinical Biochemistry, Guangdong Medical College, Zhanjiang, Guangdong 523808, P.R. China
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Pages:
4333-4340
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Published online on:
February 6, 2015
https://doi.org/10.3892/mmr.2015.3313
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Abstract
The aim of this study was to investigate the effect of the tumor‑targeting recombinant human tumor necrosis factor (rhTNF)‑α fusion protein mediated by urokinase on Sl80 tumor‑bearing mice, as well as to explore its mechanisms of action. Furthermore, the study aimed to observe the effect of the protein on liver and kidney function. rhTNF‑α fusion protein prokaryotic expression vectors were constructed using genetic engineering techniques, and were introduced into Escherichia coli. Expression of the fusion protein was induced, and it was then separated and purified in order to determine its cytotoxic activity on L929 cells. Kunming mice were randomly divided into four groups after being inoculated with S180 tumor cells. The groups were then injected with saline (control group, group S), or saline with 0.1 µg/ml fusion protein (low dose group, group L), 0.2 µg/ml fusion protein (middle dose group, group M) or 0.3 µg/ml (high dose group, group H). The mice were sacrificed after 12 days and liver [mg/kg; (liver weight/body weight) x 1,000] and kidney [mg/kg; (kidney weight/body weight) x 1,000] indices, tumor weight, the percentage reduction in mean tumor size, and the levels of alanine transaminase (ALT), albumin (ALB), creatinine (Cr) and blood urea nitrogen (BUN) in each group of mice were determined. In addition, the levels of urokinase‑type plasminogen activator (uPA), the expression of bcl‑2, bax and vascular endothelial growth factor (VEGF), and the percentage of apoptotic cells were measured with an enzyme‑linked immunosorbent assay, streptavidin‑biotin complex of immunohistochemistry and terminal deoxynucleotidyl transferase‑mediated dUTP nick end labeling, respectively. The fusion protein significantly inhibited the growth of S180 tumor cells in vivo in a dose‑dependent manner. With an increase in the dose of fusion protein, ALT, uPA, bcl‑2 and VEGF levels decreased, and ALB levels increased. However, liver and kidney indices and bax expression were not significantly altered. Cr and BUN levels did not change significantly in the low and middle dose groups, but did increase in the high dose group. Compared with the control group, the percentage of apoptotic cells in the high‑dose group was significantly higher. In conclusion, the fusion protein significantly inhibited S180 tumor growth in a mouse model, possibly by reducing the levels of uPA, bcl‑2 and VEGF. There was a mildly toxic effect on the kidneys with the high dose, but a protective effect in the liver.
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View References
|
1
|
Van Ostade X, Vandenabeele P, Everaerdt B,
et al: Human TNF mutants with selective activity on the p55
receptor. Nature. 361:266–269. 1993. View
Article : Google Scholar : PubMed/NCBI
|
|
2
|
Gerstein ES, Shcherbakov AI, Kaz’min AI,
et al: Urokinase and tissue plasminogen activators and their type-1
inhibitor (PAI-1) in gastric cancer. Vopr Onkol. 49:165–169.
2003.In Russian.
|
|
3
|
Morodomi Y, Yano T, Kinoh H, et al:
BioKnife, a uPA activity-dependent oncolytic Sendai virus,
eliminates pleural spread of malignant mesothelioma via
simultaneous stimulation of uPA expression. Mol Ther. 20:769–777.
2012. View Article : Google Scholar : PubMed/NCBI
|
|
4
|
Thummarati P, Wijitburaphat S, Prasopthum
A, et al: High level of urokinase plasminogen activator contributes
to cholangiocarcinoma invasion and metastasis. World J
Gastroenterol. 18:244–250. 2012. View Article : Google Scholar : PubMed/NCBI
|
|
5
|
Wang S, Chen T, Chen R, et al: Emodin
loaded solid lipid nanoparticles: preparation, characterization and
anti-tumor activity studies. Int J Pharm. 430:238–246. 2012.
View Article : Google Scholar : PubMed/NCBI
|
|
6
|
Zhang XJ, Yue J and Zhao TB: Expression
change of TNF-α in myocardium and hepatic tissue of rats with
compound stress of hyperthermia and lipopolysaccharide. Asian Pac J
Trop Med. 6:300–304. 2013. View Article : Google Scholar : PubMed/NCBI
|
|
7
|
Phalitakul S, Okada M, Hara Y, et al:
Vaspin prevents methylglyoxal-induced apoptosis in human vascular
endothelial cells by inhibiting reactive oxygen species generation.
Acta Physiol (Oxf). 209:212–219. 2013.
|
|
8
|
Han W, Wang S, Liang R, et al: Non-ionic
surfactant vesicles simultaneously enhance antitumor activity and
reduce the toxicity of cantharidin. Int J Nanomedicine.
8:2187–2196. 2013.PubMed/NCBI
|
|
9
|
Jiang Y, Jiang X, Law K, et al: Enhanced
anti-tumor effect of 9-nitro-camptothecin complexed by
hydroxypropyl β cyclodextrin and safety evaluation. Int J Pharm.
415:252–258. 2011. View Article : Google Scholar : PubMed/NCBI
|
|
10
|
Zhou S, Yang Y, Yang Y, et al: Combination
therapy of VEGF-trap and gemcitabine results in improved anti-tumor
efficacy in a mouse lung cancer model. PLoS One. 8:e685892013.
View Article : Google Scholar : PubMed/NCBI
|
|
11
|
Weber WA, Czernin J, Phelps ME and
Herschman HR: Technology Insight: novel imaging of molecular
targets is an emerging area crucial to the development of targeted
drugs. Nat Clin Pract Oncol. 5:44–54. 2008. View Article : Google Scholar
|
|
12
|
Klener P Jr and Klener P:
Molecularly-targeted and biological anti-cancer therapy. Folia Biol
(Praha). 58:1–6. 2012.
|
|
13
|
McGuire AT and Mangroo D: Cex1p is a novel
cytoplasmic component of the Saccharomyces cerevisiae nuclear tRNA
export machinery. EMBO J. 26:288–300. 2007. View Article : Google Scholar : PubMed/NCBI
|
|
14
|
Zhao Q, Hou G, Huang D and Chen S:
Construction and expression of hTNF-alpha fusion protein mediated
by MMP1. Sheng WuYi Xue Gong Cheng Xue Za Zhi. 28:534–537. 2011.In
Chinese.
|
|
15
|
Jia FL and Khan A: Studies on acute
toxicity and serum level changes of tumor necrosis factor. Zhongguo
Yi Xue Ke Xue Yuan Xue Buo. 11:302–305. 1989.In Chinese.
|
|
16
|
Horn LC, Pippig S, Raptis G, et al:
Clinical relevance of urokinase type plasminogen activator and its
inhibitor type 1 (PAI-1) in squamous cell carainoma of the uterine
cervix. Aust N Z J Obstet Gynaecol. 42:383–386. 2002. View Article : Google Scholar : PubMed/NCBI
|
|
17
|
Strasser A, Cory S and Adams JM:
Deciphering the rules of programmed cell death to improve therapy
of cancer and other diseases. EMBO J. 30:3667–3683. 2011.
View Article : Google Scholar : PubMed/NCBI
|
|
18
|
Sivaprasad S, Govardhan B, Harithakrishna
R, et al: Association of vascular endothelial growth factor (VEGF)
gene polymorphism and increased serum VEGF concentration with
pancreatic adenocarcinoma. Pancreatology. 13:267–272. 2013.
View Article : Google Scholar : PubMed/NCBI
|
|
19
|
Gu CP and Zhang YP: Progress on anti-tumor
mechanism of tumor necrosis factor-alpha. Bulletin of Chinese
Cancer. 16:102–105. 2007.In Chinese.
|
