重组人血管内皮抑制素促进肺癌细胞共培养体系中血管内皮细胞的凋亡机制研究

摘要

目的研究重组人血管内皮抑制素注射液(Endostar,恩度)促进与肺癌细胞A549共培养体系中的人脐静脉内皮细胞(HUVEC)凋亡及其机制研究.方法通过四甲基偶氮唑蓝(MTT),评价恩度对HUVECs增殖抑制作用；以流式细胞术Annexin V-FITC测定细胞凋亡率；以免疫细胞化学法测定恩度对抑凋亡蛋白Bcl-2和促凋亡蛋白Bax的影响；以Western blotting检测恩度对Bcl-2和Bax蛋白表达的影响.结果恩度能显著抑制肺癌细胞A549/HUVEC共培养体系中HUVEC的增殖；促进HUVECs凋亡；Annexin V-FITC测定显示10、20、40、80、100μg/mL浓度的恩度的凋亡率分别为(5.29±1.21)%、(8.33±0.62)%、(14.81±0.77)%、(21.87±0.96)%、(26.15±1.06)%；免疫细胞化学法和Western blotting检测显示恩度显著下调Bcl-2和上调Bax蛋白的表达.结论结果表明恩度能通过调控Bcl-2家族蛋白促进肺癌细胞与HUVEC共培养中血管内皮细胞的凋亡.%　　Objective To study the mechanism of recombinant human endostatin injection (Endostar) on promoting the apoptosis of human umbilical vein endothelial cells (HUVEC) in non–small lung cancer cell line A549 co-culture system. Methods Tetrazolium blue (MTT) and was used for the evaluation on proliferation inhibition of Endostar in HUVECs;Annexin V-FITC flow cytometry was used to determine the apoptosis rate;the effect of Endostar on Bcl-2 and Bax proteins expression in HUVEC by immunocytochemistry and Western blotting. Results Endostar can significantly inhibit HUVEC proliferation and significantly promote its apoptosis in A549 co-cultured system;the apoptosis rate were 5.29±1.21%(10μg/mL),8.33±0.62%(20μg/mL),14.81±0.77%(40μg/mL),21.87±0.96%(80μg/mL),26.15±1.06%(100μg/mL) by Annexin V-FITC tests;Endostar significantly down-regulated Bcl-2 and up-regulated Bax proteins expression.Conclusions Endostar can promote HUVEC apoptosis in non-small lung cancer cell co-cultured system via regulating Bcl-2 family proteins expression.