PLK1抑制剂HS-10159在NSCLC中放疗增敏机制的研究

发布时间：2018-05-10 12:12

本文选题：非小细胞肺癌 + PLK1　；参考：《天津医科大学》2016年博士论文

【摘要】：目的:有丝分裂中关键步骤包含G2/M期过渡、多种细胞器的成熟、遗传信息的损伤修复等,PLK1在其中有重要的调节功能,而PLK1在哺乳动物肿瘤中高表达,并与肿瘤细胞的增殖及患者的预后密切相关,已成为研究热点。本研究拟通过临床病理资料分析PLK1在NSCLC组织中表达及与临床预后相关因素,通过细胞学实验初步探究PLK1抑制剂HS-10159联合放疗对NSCLC细胞周期、细胞凋亡的影响及蛋白表达变化,探究PLK1抑制剂HS-10159联合放疗对NSCLC放疗增敏相关分子机制。为临床靶向应用PLK1小分子抑制剂HS-10159治疗NSCLC提供理论依据。方法:1.通过免疫组织化学的科研方法检测分析并科学评估polo激酶家族PLK1蛋白在NSCLC癌旁组织、NSCLC组织以及NSCLC脑转移组织中的表达情况,运用统计学SPSS软件预测PLK1蛋白表达状态与肺癌癌旁、肺癌及肺癌脑转移患者预后相关性及与各相关临床病理学参数间的关系。2.选取三种NSCLC细胞系A549,SPC,LTEP,通过体外MTS实验,克隆形成实验探究PLK1抑制剂HS-10159对非小细胞肺癌细胞生长增殖抑制能力及克隆形成抑制能力,观察药物对NSCLC放疗增敏性作用。3.检测PLK1、P-PLK1在三种NSCLC细胞系A549,SPC,LTEP细胞中的表达,通过流式分析细胞周期、细胞凋亡实验及细胞蛋白印迹实验探究给予NSCLC细胞系单纯药物处理、单纯放疗处理及药物联合放疗处理对NSCLC细胞有丝分裂周期(主要G2/M期)、细胞凋亡的影响及PLK1、P-PLK1蛋白在不同实验组的表达变化,探究PLK1抑制剂HS-10159联合放疗对NSCLC放疗增敏的相关分子机制。结果:1.通过对44例NSCLC组织,19例NSCLC癌旁组织和41例NSCLC脑转移组织以及7例原发肺癌及自身单发脑转移对照患者组织中PLK1表达结果分析显示:PLK1蛋白在NSCLC中表达明显高于肺癌癌旁组织;在生存分析中发现:PLK1蛋白表达阴性的NSCLC患者的总生存时间显著长于PLK1蛋白阳性表达患者。PLK1在肺癌及脑转移中表达无统计学差异。2.体外细胞增殖实验显示PLK1抑制剂HS-10159能明显抑制NSCLC癌细胞的增殖能力,单纯PLK1抑制剂HS-10159药物处理LTEP,SPC及A549细胞,给药后0.5h-6h内P-PLK1表达明显受抑制,24h P-PLK1表达恢复甚至高于对照组。3.PLK1抑制剂HS-10159能抑制NSCLC肿瘤细胞克隆形成能力,对A549细胞有明显的放疗增敏性,对LTEP,SPC细胞无明显放疗增敏性。PLK1抑制剂HS-10159对不同NSCLC细胞系有不同放疗增敏性。4.三种NSCLC细胞随单次给予的射线剂量的增加,P-PLK1蛋白表达表现为应激性增高,给予单纯药物能抑制3种NSCLC P-PLK1的水平,给予药物联合放疗能抑制单纯放疗所引起的P-PLK1水平的升高。5.PLK1抑制剂HS-10159联合射线治疗明显增加NSCLC细胞有丝分裂G2/M期阻滞,较空白对照组细胞死亡及凋亡明显增多,但PLK1抑制剂HS-10159联合放疗处理组较单纯PLK1抑制剂HS-10159组细胞凋亡无明显统计学差异。结论:1.临床病例统计结果显示PLK1蛋白的异常高表达与NSCLC的预后相关,PLK1蛋白有可能作为预测NSCLC患者的预后指标。2.PLK1抑制剂HS-10159抑制NSCLC细胞增殖能力及肿瘤细胞克隆形成能力,对不同NSCLC细胞系有不同放疗增敏性。3.放疗能导致NSCLC细胞P-PLK1蛋白表达短时间内应激性增高,PLK1抑制剂HS-10159联合射线治疗组能抑制单纯射线所引起的P-PLK1蛋白表达的升高。PLK1抑制剂HS-10159联合射线治疗组明显增加NSCLC细胞有丝分裂周期G2/M期阻滞,联合治疗组较单纯药物组细胞凋亡差异无明显统计学意义。PLK1抑制剂HS-10159对NSCLC的放疗增敏机制及参与的信号通路有待进一步探究。
[Abstract]:Objective: the key steps in mitosis include the G2/M phase transition, the maturation of various organelles, the damage and repair of genetic information, and so on. PLK1 has important regulatory functions. PLK1 is highly expressed in mammalian tumors and closely related to the proliferation of tumor cells and the prognosis of the patients. This study is to be used in clinical pathology. Data analysis of the expression of PLK1 in NSCLC tissue and related factors of clinical prognosis, through cytological experiments, preliminary exploration of PLK1 inhibitor HS-10159 combined with radiotherapy on NSCLC cell cycle, the effect of apoptosis and the change of protein expression, explore the molecular mechanism of PLK1 inhibitor HS-10159 combined with radiotherapy for NSCLC radiation sensitization. PLK1 small molecule inhibitor HS-10159 provides theoretical basis for the treatment of NSCLC. Methods: 1. through the analysis of immunohistochemical method, the expression of PLK1 protein in the Polo kinase family in the adjacent tissue of NSCLC, NSCLC tissue and NSCLC brain metastases, and the prediction of the state of the expression of PLK1 protein and the lung by the system of SPSS software The relationship between the prognosis and the related clinicopathological parameters in the patients with cancer, lung and lung cancer, and the relationship with the related clinicopathological parameters.2. selected three NSCLC cell lines A549, SPC, LTEP. Through the MTS experiment in vitro, the cloning and forming experiment explored the inhibition ability of PLK1 inhibitor HS-10159 to the proliferation of non-small cell lung cancer cells and the inhibitory ability of the clone formation. The sensitizing effect of drugs on NSCLC radiotherapy.3. was used to detect PLK1, P-PLK1 was expressed in three NSCLC cell lines A549, SPC, LTEP cells. Through flow analysis of cell cycle, apoptosis experiment and cell Western blotting, the NSCLC cell line was treated with pure drug treatment. Only radiotherapy treatment and drug combined radiotherapy were used to treat NSCLC cells. The effect of PLK1, P-PLK1 protein expression changes in different experimental groups and the related molecular mechanism of PLK1 inhibitor HS-10159 combined with radiotherapy for NSCLC radiation sensitization. Results: 1.: 1. through 44 cases of NSCLC tissue, 19 cases of NSCLC para cancer tissue and 41 cases of NSCLC brain metastases and 7 cases of primary lung cancer and self The analysis of PLK1 expression in the tissue of single brain metastases showed that the expression of PLK1 protein in NSCLC was significantly higher than that of lung cancer adjacent to lung cancer; in survival analysis, the total survival time of NSCLC patients with negative PLK1 protein expression was significantly longer than that of PLK1 positive expression patients with no statistical difference in expression of.PLK1 in lung cancer and brain metastases. In vitro cell proliferation test showed that PLK1 inhibitor HS-10159 could significantly inhibit the proliferation of NSCLC cancer cells. LTEP, SPC and A549 cells were treated with PLK1 inhibitor HS-10159 drugs only. The expression of 0.5h-6h P-PLK1 was obviously inhibited after administration, 24h P-PLK1 expression was even higher than that of the control group. The clone formation ability, A549 cells have obvious radiation sensitization, LTEP, SPC cells have no obvious radiation sensitization.PLK1 inhibitor HS-10159 to different NSCLC cell lines with different radiation sensitization of.4. three NSCLC cells with the increase of single dose of radiation, P-PLK1 protein expression is increased stress, giving simple drugs can inhibit The level of 3 kinds of NSCLC P-PLK1, drug combined radiotherapy can inhibit the increase of P-PLK1 level caused by radiotherapy alone,.5.PLK1 inhibitor HS-10159 combined radiation therapy significantly increased the NSCLC cell mitotic G2/M phase block, compared with the blank control group, cell death and apoptosis increased significantly, but the PLK1 inhibitor HS-10159 combined with radiotherapy group was more than that of the control group. There is no significant difference in apoptosis in group HS-10159 of simple PLK1 inhibitor. Conclusion: 1. clinical case statistics show that the abnormal high expression of PLK1 protein is related to the prognosis of NSCLC. The PLK1 protein may be a prognostic indicator of NSCLC patients, the.2.PLK1 inhibitor HS-10159 inhibits the proliferation of NSCLC cells and the formation of tumor cell clones. Force, different NSCLC cell lines have different radiation sensitization,.3. radiation can lead to short time stress in NSCLC cell P-PLK1 protein expression, PLK1 inhibitor HS-10159 combined radiation therapy group can inhibit the increase of P-PLK1 protein expression caused by simple ray,.PLK1 inhibitor HS-10159 combined ray treatment group obviously increased NSCLC cells. There is no significant difference in apoptosis between the combined treatment group and the single drug group in the G2/M phase of mitosis period. The mechanism of.PLK1 inhibitor HS-10159 and the signaling pathway involved in NSCLC need to be further explored.

【学位授予单位】：天津医科大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R734.2

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