选择性Bcl-2抑制剂ABT-199联合卡铂对非小细胞肺癌细胞株A549的杀伤效应

发布时间：2018-04-24 06:27

本文选题：NSCLC + A549　；参考：《安徽医科大学》2015年硕士论文

【摘要】：背景与目的:肺癌是严重危害人类健康的常见恶性肿瘤,化疗是其重要治疗手段之一,然而肿瘤耐药的产生往往造成化疗疗效不佳。有研究发现抗凋亡蛋白Bcl-2家族的过表达参与了肿瘤的耐药。在铂类耐药的肺癌患者,Bcl-2也过表达。本实验研究Bcl-2抑制剂ABT-199■联合卡铂对人非小细胞肺癌细胞A549凋亡的增效作用。方法:四甲基偶氮唑蓝(MTT)法检测A BT-199和卡铂不同浓度和作用时间对A 549细胞的增殖抑制作用并计算IC2Q作为联合实验参考剂量,检测低剂量两药联合作用24 h对A 549细胞的增殖抑制作用。流式细胞术(FCM)检测ABT-199对A 5 4 9细胞周期的影响以及两种药物联合作用下细胞凋亡率的变化;Western blot法检测凋亡相关蛋白Bcl-2、Bax、Bcl-xl、Mcl-1、Caspase-3和Caspase-9的表达。结果:ABT-199和卡铂对A549细胞有增殖抑制作用且呈时间和浓度依赖性,低剂量ABT-199联合卡铂的抑制率高于卡铂单药(P0.0l)。FCM结果显示,随着ABT-199浓度的升高,A549细胞发生不同程度的Go/G!期阻滞,联合用药组细胞凋亡率(72.4±2.2)%较卡铂单药组(37.3±1.8)°/。明显增高。Western blot结果显示,同单药组相比,联合用药组细胞Bcl-2水平明显下调(P0.01),Bcl-xl、Mcl-1无明显变化,同时B ax水平增高,Caspase-3和Caspase-9活性增加(P0.01)。结论:ABT-199和卡怕均可有效抑制人非小细胞肺癌细胞系A549增殖,ABT-199可诱导细胞发生Go/G,期阻滞,对卡铂诱导A 549细胞凋亡具有增效作用。
[Abstract]:Background & objective: lung cancer is a common malignant tumor that seriously endangers human health. Chemotherapy is one of the most important treatment methods. It has been found that the overexpression of anti-apoptotic protein Bcl-2 family is involved in tumor resistance. Bcl-2 is also overexpressed in patients with platinum-resistant lung cancer. The aim of this study was to investigate the synergistic effect of Bcl-2 inhibitor ABT-199 and carboplatin on apoptosis of human non-small cell lung cancer cell line A549. Methods: the inhibitory effects of BT-199 and carboplatin on the proliferation of A549 cells were measured by tetramethyl azolium blue tTassay. The IC2Q was used as the reference dose of the combined experiment, and the inhibitory effects of IC2Q and carboplatin on the proliferation of A549 cells were calculated. The inhibitory effect of low dose two drugs on proliferation of A549 cells for 24 h was detected. Flow cytometry (FCM) was used to detect the effect of ABT-199 on the cell cycle of A549 cells and the change of apoptosis rate under the combination of two drugs. The expression of apoptosis-related protein Bcl-2Bax-xlmcl-1mcl-1caspase-3 and Caspase-9 was detected by Western blot assay. Results the inhibition rate of low dose ABT-199 combined with carboplatin on A549 cells was higher than that of carboplatin single drug P0. 0 l).FCM. The results showed that with the increase of ABT-199 concentration, there were different degrees of Gor / GG in A549 cells. The apoptosis rate was 72.4 卤2.2% in combination group and 37.3 卤1.8 掳/ r in single carboplatin group. The results of Western blot showed that compared with the single drug group, the level of Bcl-2 in the combined treatment group was significantly decreased, while the activity of Caspase-3 and Caspase-9 was increased in the combined treatment group (P 0.01). Conclusion both 1: ABT-199 and Cabernet can effectively inhibit the proliferation of human non-small cell lung cancer cell line A549. ABT-199 can induce Go-G and phase arrest of A549 cells, which has a synergistic effect on the apoptosis of A549 cells induced by carboplatin.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R734.2

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