Effects of the C‑terminal of endostatin on the tumorigenic potential of H22 cells

Qi,Xia; Liu,Yuejian; Wei,Wei; Huang,Xiaohua; Zuo,Yunfei

doi:10.3892/br.2013.147

Effects of the C‑terminal of endostatin on the tumorigenic potential of H22 cells

Authors:
- Xia Qi
- Yuejian Liu
- Wei Wei
- Xiaohua Huang
- Yunfei Zuo
View Affiliations

Affiliations: Department of Clinical Biochemistry, Dalian Medical University, Dalian, Liaoning 116044, P.R. China, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, P.R. China
Published online on: July 22, 2013 https://doi.org/10.3892/br.2013.147
Pages: 761-765

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Endostatin is an endogenous angiogenesis inhibitor whose specific functional site has not yet been determined. In the present experiment, 13 amino acids (LCIENSFMTSFSK) were selectively deleted from the C‑terminal of endostatin and the resulting mutant endostatin was named EM13. To determine the effect of the C‑terminal deletion on the biological activity of endostatin, EM13, wild‑type endostatin and empty plasmid were transfected into H22 cells. After 48 h, the three types of transfected cells were harvested and injected into nude mice. The results demonstrated that there was no significant difference in tumor size, as determined by hematoxylin and eosin staining, between the EM13‑transfected group and the endostatin and empty plasmid groups, although the nude mice that were injected with EM13‑transfected H22 cells exhibited smaller tumors and lower density of blood vessels compared to those injected with endostatin- and empty plasmid‑transfected H22 cells. The results suggested that the 13 amino acids of the C‑terminal of endostatin do not play an important role in the tumorigenic potential of H22 cells. this experiment was unsuccessful in reproducing the results of several investigators. Therefore, the mechanism underlying the tumorigenesis of H22 cells remains to be elucidated.

View Figures

View References

1	Boehm T, Folkman J, Browder T and O’Reilly MS: Antiangiogenic therapy of experimental cancer does not induce acquired drug resistance. Nature. 390:404–407. 1997. View Article : Google Scholar
2	Li J, Dong X, Xu Z, Jiang X, Jiang H, Krissansen GW and Sun X: Endostatin gene therapy enhances the efficacy of paclitaxel to suppress breast cancers and metastases in mice. J Biomed Sci. 15:99–109. 2008. View Article : Google Scholar : PubMed/NCBI
3	Herbst RS, Hess KR, Tran HT, Tseng JE, Mullani NA, Charnsangavej C, Madden T, Davis DW, McConkey DJ, O’Reilly MS, Ellis LM, Pluda J, Hong WK and Abbruzzese JL: Phase I study of recombinant human endostatin in patients with advanced solid tumors. J Clin Oncol. 20:3792–3803. 2002. View Article : Google Scholar : PubMed/NCBI
4	Felbor U, Dreier L, Bryant RA, Ploegh HL, Olsen BR and Mothes W: Secreted cathepsin L generates endostatin from collagen XVIII. EMBO J. 19:1187–1194. 2000. View Article : Google Scholar : PubMed/NCBI
5	Lin HC, Chang JH, Jain S, Gabison EE, Kure T, Kato T, Fukai N and Azar DT: Matrilysin cleavage of corneal collagen type XVIII NC1 domain and generation of a 28-kDa fragment. Invest Ophthalmol Vis Sci. 42:2517–2524. 2001.PubMed/NCBI
6	Wen W, Moses MA, Wiederschain D, Arbiser JL and Folkman J: The generation of endostatin is mediated by elastase. Cancer Res. 59:6052–6056. 1999.PubMed/NCBI
7	Ferreras M, Felbor U, Lenhard T, Olsen BR and Delaisse J: Generation and degradation of human endostatin proteins by various proteinases. FEBS Lett. 486:247–251. 2000. View Article : Google Scholar : PubMed/NCBI
8	Sudhakar A, Sugimoto H, Yang C, Lively J, Zeisberg M and Kalluri R: Human tumstatin and human endostatin exhibit distinct antiangiogenic activities mediated by αvβ3 and α5β1 integrins. Proc Natl Acad Sci USA. 100:4766–4771. 2003.PubMed/NCBI
9	Karumanchi SA, Jha V, Ramchandran R, Karihaloo A, Tsiokas L, Chan B, Dhanabal M, Hanai JI, Venkataraman G, Shriver Z, Keiser N, Kalluri R, Zeng H, Mukhopadhyay D, Chen RL, Lander AD, Hagihara K, Yamaguchi Y, Sasisekharan R, Cantley L and Sukhatme VP: Cell surface glypicans are low-affinity endostatin receptors. Mol Cell. 7:811–822. 2001. View Article : Google Scholar : PubMed/NCBI
10	Ling Y, Yang Y, Lu N, You QD, Wang S, Gao Y, Chen Y and Guo QL: Endostar, a novel recombinant human endostatin, exerts antiangiogenic effect via blocking VEGF-induced tyrosine phosphorylation of KDR/Flk-1 of endothelial cells. Biochem Biophys Res Commun. 361:79–84. 2007. View Article : Google Scholar
11	Kim YM, Hwang S, Kim YM, Pyun BJ, Kim TY, Lee ST, Gho YS and Kwon YG: Endostatin blocks vascular endothelial growth factor-mediated signaling via direct interaction with KDR/Flk-1. J Biol Chem. 277:27872–27879. 2002. View Article : Google Scholar : PubMed/NCBI
12	Kim YM, Jang JW, Lee OH, Yeon J, Choi EY, Kim KW, Lee ST and Kwon YG: Endostatin inhibits endothelial and tumor cellular invasion by blocking the activation and catalytic activity of matrix metalloproteinase. Cancer Res. 60:5410–5413. 2000.PubMed/NCBI
13	Hanai J, Dhanabal M, Karumanchi SA, Albanese C, Waterman M, Chan B, Ramchandran R, Pestell R and Sukhatme VP: Endostatin causes G1 arrest of endothelial cells through inhibition of cyclin D1. J Biol Chem. 277:16464–16469. 2002. View Article : Google Scholar : PubMed/NCBI
14	Shichiri M and Hirata Y: Antiangiogenesis signals by endostatin. FASEB J. 15:1044–1053. 2001. View Article : Google Scholar : PubMed/NCBI
15	Hebbar M, Peyrat JP, Hornez L, Hatron PY, Hachulla E and Devulder B: Increased concentrations of the circulating angiogenesis inhibitor endostatin in patients with systemic sclerosis. Arthritis Rheum. 43:889–893. 2000. View Article : Google Scholar : PubMed/NCBI
16	Funatsu H, Yamashita H, Noma H, Shimizu E, Yamashita T and Hori S: Stimulation and inhibition of angiogenesis in diabetic retinopathy. Jpn J Ophthalmol. 45:577–584. 2001. View Article : Google Scholar : PubMed/NCBI
17	Ma L, del Soldato P and Wallace JL: Divergent effects of new cyclooxygenase inhibitors on gastric ulcer healing: Shifting the angiogenic balance. Proc Natl Acad Sci USA. 99:13243–13247. 2002. View Article : Google Scholar : PubMed/NCBI
18	Hajitou A, Grignet C, Devy L, Berndt S, Blacher S, Deroanne CF, Bajou K, Fong T, Chiang Y, Foidart JM and Noël A: The antitumoral effect of endostatin and angiostatin is associated with a down-regulation of vascular endothelial growth factor expression in tumor cells. FASEB J. 16:1802–1804. 2002.PubMed/NCBI
19	Maione TE, Gray GS, Petro J, Hunt AJ, Donner AL, Bauer SI, Carson HF and Sharpe RJ: Inhibition of angiogenesis by recombinant human platelet factor-4 and related peptides. Science. 247:77–79. 1990. View Article : Google Scholar : PubMed/NCBI
20	Vogel T, Guo N, Krutzsch HC, Blake DA, Hartmann J, Mendelovitz S, Panet A and Roberts DD: Modulation of endothelial cell proliferation, adhesion and motility by recombinant heparin-binding domain and synthetic peptides from the type I repeats of thrombospondin. J Cell Biochem. 53:74–84. 1993. View Article : Google Scholar : PubMed/NCBI
21	O’Reilly MS, Holmgren L, Shing Y, Chen C, Rosenthal RA, Moses M, Lane WS, Chao Y, Sage EH and Folkman J: Angiostatin: a novel angiogenesis inhibitor that mediates the suppression of metastases by a Lewis lung carcinoma. Cell. 79:315–328. 1994.PubMed/NCBI
22	O’Reilly MS, Boehm T, Shing Y, Fukai N, Vasois G, Lane WS, Flynn E, Birkhead JR, Olsen BR and Folkman J: Endostatin: an endogenous inhibitor of angiogenesis and tumor growth. Cell. 88:277–285. 1997.
23	Good DJ, Polverini PJ, Rastinejad F, Le Beau MM, Lemons RS, Frazier WA and Bouck NP: A tumor suppressor-dependent inhibitor of angiogenesis is immunologically and functionally indistinguishable from a fragment of thrombospondin. Proc Natl Acad Sci USA. 87:6624–6628. 1990. View Article : Google Scholar : PubMed/NCBI
24	Tolsma SS, Volpert OV, Good DJ, Frazier WA, Polverini PJ and Bouck N: Peptides derived from two separate domains of the matrix protein thrombospondin-1 have anti-angiogenic activity. J Cell Biol. 122:497–511. 1993. View Article : Google Scholar : PubMed/NCBI
25	Dawson DW, Pearce SF, Zhong R, Silverstein RL, Frazier WA and Bouck NP: CD36 mediates the in vitro inhibitory effects of thrombospondin-1 on endothelial cells. J Cell Biol. 138:707–717. 1997. View Article : Google Scholar : PubMed/NCBI
26	Cao Y, Ji RW, Davidson D, Schaller J, Marti D, Söhndel S, McCance SG, O’ Reilly M, Llinás M and Folkman J: Kringle domains of human angiostatin. Characterization of the anti-proliferative activity on endothelial cells. J Biol Chem. 271:29461–29467. 1996. View Article : Google Scholar : PubMed/NCBI
27	Dhanabal M, Ramchandran R, Volk R, et al: Endostatin: yeast production, mutants, and antitumor effect in renal cell carcinoma. Cancer Res. 59:189–197. 1999.PubMed/NCBI
28	Peroulis I, Jonas N and Saleh M: Antiangiogenic activity of endostatin inhibits C6 glioma growth. Int J Cancer. 97:839–845. 2002. View Article : Google Scholar : PubMed/NCBI
29	Yokoyama Y, Sedgewich G and Ramakrishnan S: Endostatin binding to ovarian cancer cells inhibits peritoneal attachment and dissemination. Cancer Res. 67:10813–10822. 2007. View Article : Google Scholar : PubMed/NCBI
30	Liu F, Tan G, Li J, et al: Gene transfer of endostatin enhances the efficacy of doxorubicin to suppress human hepatocellular carcinomas in mice. Cancer Sci. 98:1381–1387. 2007. View Article : Google Scholar : PubMed/NCBI
31	Hoffmann S, Wunderlich A, Celik I, et al: Paneling human thyroid cancer cell lines for candidate proteins for targeted antiangiogenic therapy. J Cell Biochem. 98:954–965. 2006. View Article : Google Scholar : PubMed/NCBI