目的
測定攜帶小鼠內皮抑素(
mES
)基因的腺病毒(
Ad-mES
)在腫瘤細胞中的表達,觀察
mES
對體外培養的血管內皮細胞的抑制作用。方法 分別以0.01、0.1、1、10及100共5個不同感染復數(MOI)的Ad-mES轉染Lewis肺癌細胞株(LLC)。采用免疫組織化學法及Western blot檢測Ad-mES轉染LLC 48 h后mES蛋白的表達;應用四甲基偶氮唑藍比色(MTT)分析法測定不同MOI值Ad-mES轉染上清對基礎狀態下及經堿性成纖維細胞生長因子(bFGF)刺激的人臍靜脈內皮細胞(ECV304)的增殖抑制作用。結果 Ad-mES轉染LLC 48 h后,LLC細胞漿內可見棕黃色mES陽性表達顆粒,培養上清可表達位于相對分子質量20 000的mES陽性條帶,而未轉染對照組呈陰性。隨轉染Ad-mES MOI值的增高,培養上清基礎條件下及經bFGF刺激的內皮細胞的抑制作用越顯著(與基礎條件下比較,P lt;0.05)。結論 構建的Ad-mES轉染LLC后能夠表達分泌mES;表達分泌于培養上清中的mES對血管內皮細胞(尤其是bFGF刺激)具有明顯的增殖抑制作用。
引用本文:
馬凌云 ,李強,劉忠令. 腺病毒介導的鼠內皮抑素基因轉染肺癌細胞體外抑制血管內皮細胞活性的實驗研究. 中國呼吸與危重監護雜志, 2008, 08(3): 214-217. doi:
復制
1. |
Folk man J.What is the evidence that tumors are angiogenesis dependent? J Natl Cancer Inst,1990,82:4-6.
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2. |
Cao Y.Endogenous angiogenesis inhibitors and their therapeutic implications.Int J Biochem Cell Biol,2001,33:357-369.
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高雪軍,伍志堅,吳小兵,等.腺相關病毒載體介導的人內皮抑素的體外表達及對內皮細胞抑制作用的研究.病毒學報,2002,18:235-239.
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顏浩,徐淑暉,周黎強,等.血管內皮生長因子及其受體在肺癌組織中表達的研究.中國呼吸與危重監護雜志,2005,4:189-192.
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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.
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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.
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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. 高雪軍,伍志堅,吳小兵,等.腺相關病毒載體介導的人內皮抑素的體外表達及對內皮細胞抑制作用的研究.病毒學報,2002,18:235-239.
- 4. 顏浩,徐淑暉,周黎強,等.血管內皮生長因子及其受體在肺癌組織中表達的研究.中國呼吸與危重監護雜志,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.