目的 測(cè)定攜帶小鼠內(nèi)皮抑素( mES )基因的腺病毒( Ad-mES )在腫瘤細(xì)胞中的表達(dá),觀察 mES 對(duì)體外培養(yǎng)的血管內(nèi)皮細(xì)胞的抑制作用。方法 分別以0.01、0.1、1、10及100共5個(gè)不同感染復(fù)數(shù)(MOI)的Ad-mES轉(zhuǎn)染Lewis肺癌細(xì)胞株(LLC)。采用免疫組織化學(xué)法及Western blot檢測(cè)Ad-mES轉(zhuǎn)染LLC 48 h后mES蛋白的表達(dá);應(yīng)用四甲基偶氮唑藍(lán)比色(MTT)分析法測(cè)定不同MOI值A(chǔ)d-mES轉(zhuǎn)染上清對(duì)基礎(chǔ)狀態(tài)下及經(jīng)堿性成纖維細(xì)胞生長(zhǎng)因子(bFGF)刺激的人臍靜脈內(nèi)皮細(xì)胞(ECV304)的增殖抑制作用。結(jié)果 Ad-mES轉(zhuǎn)染LLC 48 h后,LLC細(xì)胞漿內(nèi)可見(jiàn)棕黃色mES陽(yáng)性表達(dá)顆粒,培養(yǎng)上清可表達(dá)位于相對(duì)分子質(zhì)量20 000的mES陽(yáng)性條帶,而未轉(zhuǎn)染對(duì)照組呈陰性。隨轉(zhuǎn)染Ad-mES MOI值的增高,培養(yǎng)上清基礎(chǔ)條件下及經(jīng)bFGF刺激的內(nèi)皮細(xì)胞的抑制作用越顯著(與基礎(chǔ)條件下比較,P lt;0.05)。結(jié)論 構(gòu)建的Ad-mES轉(zhuǎn)染LLC后能夠表達(dá)分泌mES;表達(dá)分泌于培養(yǎng)上清中的mES對(duì)血管內(nèi)皮細(xì)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(qiáng),劉忠令. 腺病毒介導(dǎo)的鼠內(nèi)皮抑素基因轉(zhuǎn)染肺癌細(xì)胞體外抑制血管內(nèi)皮細(xì)胞活性的實(shí)驗(yàn)研究. 中國(guó)呼吸與危重監(jiān)護(hù)雜志, 2008, 08(3): 214-217. doi: 復(fù)制
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- 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(jiān),吳小兵,等.腺相關(guān)病毒載體介導(dǎo)的人內(nèi)皮抑素的體外表達(dá)及對(duì)內(nèi)皮細(xì)胞抑制作用的研究.病毒學(xué)報(bào),2002,18:235-239.
- 4. 顏浩,徐淑暉,周黎強(qiáng),等.血管內(nèi)皮生長(zhǎng)因子及其受體在肺癌組織中表達(dá)的研究.中國(guó)呼吸與危重監(jiān)護(hù)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.