机构地区:[1]暨南大学附属第一医院,广东广州510630 [2]暨南大学生命科技学院抗体工程中心,广东广州510630
出 处:《中国肿瘤生物治疗杂志》2016年第6期783-788,共6页Chinese Journal of Cancer Biotherapy
基 金:国家自然科学基金资助项目(No.81273814);广东省新药创制重大科技专项资助(No.2013A022100031)~~
摘 要:目的:研究碱性成纤维细胞生长因子单克隆抗体(monoclonal antibody to basic fibroblast growth factor,b FGF m Ab)联合洛铂(lobaplatin,LBP)对肺癌A549细胞增殖和凋亡的影响及其可能机制。方法:CCK-8法检测药物对A549细胞活性的影响。Annexin V-FITC/PI检测细胞凋亡率。激光共聚焦显微镜观察细胞核的形态。Western blotting检测细胞内凋亡相关蛋白表达。结果:CCK-8法检测结果显示,细胞培养48 h后,bFGF mAb组和LBP组的细胞增殖受到抑制,但两药联用后的增殖抑制作用明显高于单用LBP及bFGF mAb(P<0.05)。Annexin V-FITC/PI结果显示,联合组早期、晚期凋亡率分别为(27.83±1.23)%和(39.32±1.01)%,与b FGF mAb的(6.25±0.19)%、(14.54±0.25)%和LBP的(16.54±0.39)%、(21.02±0.98)%相比,明显高于单药组的抑制率(P<0.01)。激光共聚焦显微镜显示联合用药诱导的细胞核裂解率较单用b FGF m Ab及LBP的细胞核裂解率明显增多。Western blotting检测结果显示联合组BAX、Caspase-3、Caspase-9、PARP表达量较药物单用组明显增加,而Bcl-2、cleaved-Caspase-3、cleaved-Caspase-9、cleaved-PARP表达量明显降低。结论:b FGF m Ab联合LBP通过抑制Bcl-2、cleaved-Caspase-3、cleaved-Caspase-9、cleaved-PARP表达,上调BAX、Caspase-3、Caspase-9和PARP蛋白表达,对肺腺癌A549细胞起到抑制增殖和诱导凋亡的作用。Objective: To explore effect of monoclonal antibody to basic fibroblast growth factor (bFGF mAb) com- bined with lobaplatin (LBP) on proliferation and apoptosis of lung adenocarcinama A549 cell and their possible mecha- nisms. Methods: Effect of the drugs on viability of the A549 cell was detected by CCK8 assay. Annexin V-FITC/PI assay was used to detect apoptosis rates of the A549 cell. Laser scanning eonfocal microscope was used to observe nuclear mor- phology of the cells. Expressions of related-apoptosis proteins in the ceils were measured by Western blotting assay. Re- suits : Outcomes of CCK8 assay shown that after culturing of the cell for 48 h, proliferations of the cells in bFGF mAb and LBP groups were inhibited, but inhibited proliferation of the cell in combination of both group was obviously higher than those in single LBP and single bFGF mAb groups (P 〈 0.05). Results of Annexin V-FITC/PI assay suggested that early and later apoptosis rate in bFGFmAb combined with LBP group was (27.83 ± 1.23 ) % and (39.32 ± 1.01 ) % respec- tively, comparing with those in bFGF mAb group ( [ 6.25 ± 0.19 ] % and [ 14.54 ±0.25 ] % ) and those in LBP group( [ 16. 54 ± 0.39 ] % and [ 21.01 ± 0.98 ] % ), early and later apoptosis rates of the combination group was significantly higher than those of the single drug gYoups (P 〈 0.01 ). The result of laser scanning confocal microscope showed that cyto- nuclear fragmentation rate of the combination drugs group was more significantly increased than those of the single LBP and the single bFGF mAb groups. Western blotting assay displayed that in the combination group, expressions of BAX, Caspase-3, Caspase-9 and PARPs proteins obviously increased and expressions of Bcl-2, cleaved-Caspase-3, cleaved- Caspase-9 and cleaved-PARP proteins obviously decreased as comparing with those in both of the single drug groups. Con- clusion : bFGF mAb combined with LBP could inhibit proliferation of lung adenocarcinama A549 cell and induce apopt
关 键 词:碱性成纤维细胞生长因子单克隆抗体 洛铂 肺腺癌 细胞增殖 凋亡
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