Mouse endostatin gene transfected lung cancer cells inhibit proliferation of endothelial cells in vitro_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

MA Lingyun ¹ ,  LI Qiang ² , LIU Zhongling ²

Department of Respiratory Medicine，The 1st Affiliated Hospital，General Hospital of PLA.Beijing，100037，China Corresponding author：LI Qiang，E-mail：Liqres@sh163.net;

Corresponding author：

LI Qiang, Email: Liqres@sh163.net

Keywords：

Endostatin; Adenovirus; Gene transfection; Mouse; Basic fibroblast growth factor

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Objective To observe the expression of adenovirus vector coding for mouse endostatin gene（Ad-mES） in lung cancer cells and its antiangiogenic activity in human umbilical vein endothelial cells（ECV304） in vitro.Methods Lewis lung cancer（LLC） cells were transfected with Ad-mES at different multiplicity of infection（MOI）.The expression of mES in LLC cells and supernatant after 48 hours was detected by immunohistochemical staining and Western blot respectively.The inhibitory effect of supernatant at different MOI on ECV304 non-stamulated and stimulated by basic fibroblast growth factor（bFGF） was measured by methyl thiazolyl tetrazolium（MTT） assay.Results After transfected for 48 hours，endostatin was identified in the cell plasma of infected LLC and negative result was founded in non-infected LLC.Western blot revealed band of endostatin in 20 kDa in culture supernatant of infected LLC and negative results in non-infected LLC.The inhibitory effects on ECV304 cell proliferation were ber at higher MOI，and the difference was significant between stimulated and non-stamulated cells by bFGF（P lt;0.05）.Conclusion Ad-mES can transfect and express endostatin effectively in LLC with biological activity

Citation： MA Lingyun,LI Qiang,LIU Zhongling. Mouse endostatin gene transfected lung cancer cells inhibit proliferation of endothelial cells in vitro. Chinese Journal of Respiratory and Critical Care Medicine, 2008, 08(3): 214-217. doi: Copy

1.	Folk man J.What is the evidence that tumors are angiogenesis dependent? J Natl Cancer Inst，1990，82：4-6.
2.	Cao Y.Endogenous angiogenesis inhibitors and their therapeutic implications.Int J Biochem Cell Biol，2001，33：357-369.
3.	高雪军，伍志坚，吴小兵，等.腺相关病毒载体介导的人内皮抑素的体外表达及对内皮细胞抑制作用的研究.病毒学报，2002，18：235-239.
4.	颜浩，徐淑晖，周黎强，等.血管内皮生长因子及其受体在肺癌组织中表达的研究.中国呼吸与危重监护杂志，2005，4：189-192.
5.	Taddei L，Chiarugi P，Brogelli L，et al.Inhibitory effect of fμll-length human endostatin on in vitro angiogenesis.Biochem Biophys Res Commun，1999，263：340-345.
6.	Hohenenster E，Sasaki T，Olsen B，et al.Crystal structure of the angiogenesis inhibitor endostatin at 1.5A resolution.EMBO J，1998，17：1656-1664.
7.	Lee SJ，Jang JW，Kim YM，et al.Endostatin binds to the catalytic domain of matrix metalloproteinase-2.FEBS Lett，2002，519：147-152.
8.	Kim YM，Jang JW，Lee OH，et al.Endostatin inhibits endothelial and tumor cellar invasionbuy blocking the activation and catalytic activity of matrix metalloproteinase-2.Cancer Res，2000，60：5410-5413.
9.	MacDonald NJ，Shivers WY，Narum DL，et al.Endostatin binds trypomyosin.A potential modulator of the antirumor activity of endostatin.J Biol Chem，2001，276：25190-25196.
10.	Dhanabal M，Ramchandran R，Waterman MJF，et al.Endostatin induces endothelial cell apoptosis.J Biol Chem，1999，274：11721-11726.
11.	Shichiri M，Hirata Y.Antiangiogenesis signals by endostatin.FASEB J，2001，15：1044-1053.
12.	Kang HY，Shim D，Kang SS，et al.Protein kinase B inhibits endostatin-induced apoptosis in HUVECs.J Biochem Mol Biol，2006，39：97-104.

1. Folk man J.What is the evidence that tumors are angiogenesis dependent? J Natl Cancer Inst，1990，82：4-6.
2. Cao Y.Endogenous angiogenesis inhibitors and their therapeutic implications.Int J Biochem Cell Biol，2001，33：357-369.
3. 高雪军，伍志坚，吴小兵，等.腺相关病毒载体介导的人内皮抑素的体外表达及对内皮细胞抑制作用的研究.病毒学报，2002，18：235-239.
4. 颜浩，徐淑晖，周黎强，等.血管内皮生长因子及其受体在肺癌组织中表达的研究.中国呼吸与危重监护杂志，2005，4：189-192.
5. Taddei L，Chiarugi P，Brogelli L，et al.Inhibitory effect of fμll-length human endostatin on in vitro angiogenesis.Biochem Biophys Res Commun，1999，263：340-345.
6. Hohenenster E，Sasaki T，Olsen B，et al.Crystal structure of the angiogenesis inhibitor endostatin at 1.5A resolution.EMBO J，1998，17：1656-1664.
7. Lee SJ，Jang JW，Kim YM，et al.Endostatin binds to the catalytic domain of matrix metalloproteinase-2.FEBS Lett，2002，519：147-152.
8. Kim YM，Jang JW，Lee OH，et al.Endostatin inhibits endothelial and tumor cellar invasionbuy blocking the activation and catalytic activity of matrix metalloproteinase-2.Cancer Res，2000，60：5410-5413.
9. MacDonald NJ，Shivers WY，Narum DL，et al.Endostatin binds trypomyosin.A potential modulator of the antirumor activity of endostatin.J Biol Chem，2001，276：25190-25196.
10. Dhanabal M，Ramchandran R，Waterman MJF，et al.Endostatin induces endothelial cell apoptosis.J Biol Chem，1999，274：11721-11726.
11. Shichiri M，Hirata Y.Antiangiogenesis signals by endostatin.FASEB J，2001，15：1044-1053.
12. Kang HY，Shim D，Kang SS，et al.Protein kinase B inhibits endostatin-induced apoptosis in HUVECs.J Biochem Mol Biol，2006，39：97-104.

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