光动力疗法联合顺铂对顺铂耐药肿瘤细胞的体外治疗效应

发布时间：2018-03-27 23:41

本文选题：光动力治疗　切入点：顺铂　出处：《重庆医科大学》2017年硕士论文

【摘要】：背景:铂类抗肿瘤药物是很多实体肿瘤的一线化疗药物,耐药性的产生是导致肿瘤化疗失败的重要原因。因此,需要一些新的策略来克服顺铂耐药。光动力疗法(Photodynamic therapy,PDT)是一种获得临床认可的癌症治疗方法,基于光敏剂、光照和氧气之间的光化学反应。光动力疗法分两个阶段完成,先给予光敏剂处理,随后再用适当波长的光照射局部肿瘤激活光敏剂,从而发挥抗肿瘤效应。将多种具有不同毒性作用的治疗方法联合应用,是当代肿瘤医学提高治疗疗效常用的策略。联合治疗时,由一种治疗方法诱导的抗性有可能会被另一种方法克服。目的:本文将探讨光敏剂焦脱镁叶绿酸甲酯介导的光动力治疗对顺铂耐药肺癌细胞的灭活,及PDT与顺铂(DDP)的联合效应。方法:采用肺癌细胞A549及其DDP耐药株A549/DDP。实验分为对照组、DDP组、PDT组和PDT+DDP组。对照组不加药物,DDP组给予14μmol/L DDP处理,PDT组给予2μmol/L MPPa、2.4 J/cm2光照处理,PDT+DDP组联合应用PDT和DDP。以CCK-8法测定细胞活性,应用Annexin V-FITC/PI双染检测细胞凋亡;2',7'-二氯荧光黄双乙酸盐(DCFH-DA)检测细胞内活性氧(ROS)水平;蛋白印迹法检测Caspase-3、Bcl-2和Bax蛋白表达。结果:PDT在A549和A549/DDP细胞产生毒性,与对照组相比存活率降低(P0.05);与对照组、DDP组和PDT组相比,在二株细胞中PDT+DDP组的存活率最低(P0.05),定量分析联合指数分别为1.11、1.10,提示联合效应均为相加。在二株细胞中,PDT+DDP组凋亡率和细胞内ROS均高于对照组、DDP组及PDT组(P0.05)。蛋白质印迹法检测发现在二株细胞,与对照组相比,PDT组和PDT+DDP组,Caspase-3和Bax蛋白表达升高,Bcl-2蛋白表达降低(P0.05)。结论:PDT或联合DDP对耐药肺癌细胞具有明显杀伤效应,主要通过ROS?线粒体途径凋亡实现;PDT与DDP的联用效应为相加。
[Abstract]:Background: platinum antitumor drugs are the first-line chemotherapeutic agents in many solid tumors, and the occurrence of drug resistance is an important factor leading to the failure of chemotherapy. New strategies are needed to overcome cisplatin resistance. Photodynamic therapy (PDT) is a clinically recognized cancer treatment based on photochemical reactions between Guang Min, light and oxygen. Photodynamic therapy is done in two phases. First, Guang Min was given, and then local tumor was irradiated with appropriate wavelength of light to activate Guang Min, so as to exert anti-tumor effect. A variety of treatment methods with different toxic effects were used in combination. It is a commonly used strategy in modern oncology medicine to improve the therapeutic effect. Resistance induced by one therapeutic method may be overcome by another. Objective: to investigate the inactivation of cisplatin resistant lung cancer cells induced by photodynamic therapy mediated by methyl pyrodeoxylate, a Guang Min agent, in vitro. Methods: lung cancer cell line A549 and its DDP resistant strain A549 / DDP were divided into control group and PDT DDP group. The control group was treated with 14 渭 mol/L DDP and the control group was given 2 渭 mol/L MPPA 2.4 J/cm2 illumination. PDT and DDP were used in the DDP group. The cell activity was measured by CCK-8 assay. Annexin V-FITC/PI double staining was used to detect the expression of Caspase-3 Bcl-2 and Bax protein in A549 and A549/DDP cells, and DCFH-DA-DCFH-DAA was used to detect the expression of Caspase-3 Bcl-2 and Bax protein in A549 and A549/DDP cells. Compared with the control group, the survival rate was lower than that in the control group, and compared with the control group and the PDT group, the survival rate was lower than that in the control group. The survival rate of PDT DDP group was the lowest, and the quantitative analysis index was 1.111.10, indicating that the combination effect was additive. The apoptosis rate and intracellular ROS of PDT DDP group were higher than those of control group and PDT group. Two cell lines were detected by qualitative blotting. Compared with the control group and PDT DDP group, the expression of Caspase-3 and Bax protein increased and the expression of Bcl-2 protein decreased P0.05.Conclusion the effect of PDT / PDT or combined DDP on drug-resistant lung cancer cells is obvious. Mitochondrial pathway apoptosis can achieve the combined effect of PDT and DDP.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R730.5

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