本文選題：非小細胞肺癌 + PLK1��； 參考：《天津醫(yī)科大學(xué)》2016年博士論文

【摘要】：目的:有絲分裂中關(guān)鍵步驟包含G2/M期過渡、多種細胞器的成熟、遺傳信息的損傷修復(fù)等,PLK1在其中有重要的調(diào)節(jié)功能,而PLK1在哺乳動物腫瘤中高表達,并與腫瘤細胞的增殖及患者的預(yù)后密切相關(guān),已成為研究熱點。本研究擬通過臨床病理資料分析PLK1在NSCLC組織中表達及與臨床預(yù)后相關(guān)因素,通過細胞學(xué)實驗初步探究PLK1抑制劑HS-10159聯(lián)合放療對NSCLC細胞周期、細胞凋亡的影響及蛋白表達變化,探究PLK1抑制劑HS-10159聯(lián)合放療對NSCLC放療增敏相關(guān)分子機制。為臨床靶向應(yīng)用PLK1小分子抑制劑HS-10159治療NSCLC提供理論依據(jù)。方法:1.通過免疫組織化學(xué)的科研方法檢測分析并科學(xué)評估polo激酶家族PLK1蛋白在NSCLC癌旁組織、NSCLC組織以及NSCLC腦轉(zhuǎn)移組織中的表達情況,運用統(tǒng)計學(xué)SPSS軟件預(yù)測PLK1蛋白表達狀態(tài)與肺癌癌旁、肺癌及肺癌腦轉(zhuǎn)移患者預(yù)后相關(guān)性及與各相關(guān)臨床病理學(xué)參數(shù)間的關(guān)系。2.選取三種NSCLC細胞系A(chǔ)549,SPC,LTEP,通過體外MTS實驗,克隆形成實驗探究PLK1抑制劑HS-10159對非小細胞肺癌細胞生長增殖抑制能力及克隆形成抑制能力,觀察藥物對NSCLC放療增敏性作用。3.檢測PLK1、P-PLK1在三種NSCLC細胞系A(chǔ)549,SPC,LTEP細胞中的表達,通過流式分析細胞周期、細胞凋亡實驗及細胞蛋白印跡實驗探究給予NSCLC細胞系單純藥物處理、單純放療處理及藥物聯(lián)合放療處理對NSCLC細胞有絲分裂周期(主要G2/M期)、細胞凋亡的影響及PLK1、P-PLK1蛋白在不同實驗組的表達變化,探究PLK1抑制劑HS-10159聯(lián)合放療對NSCLC放療增敏的相關(guān)分子機制。結(jié)果:1.通過對44例NSCLC組織,19例NSCLC癌旁組織和41例NSCLC腦轉(zhuǎn)移組織以及7例原發(fā)肺癌及自身單發(fā)腦轉(zhuǎn)移對照患者組織中PLK1表達結(jié)果分析顯示:PLK1蛋白在NSCLC中表達明顯高于肺癌癌旁組織;在生存分析中發(fā)現(xiàn):PLK1蛋白表達陰性的NSCLC患者的總生存時間顯著長于PLK1蛋白陽性表達患者。PLK1在肺癌及腦轉(zhuǎn)移中表達無統(tǒng)計學(xué)差異。2.體外細胞增殖實驗顯示PLK1抑制劑HS-10159能明顯抑制NSCLC癌細胞的增殖能力,單純PLK1抑制劑HS-10159藥物處理LTEP,SPC及A549細胞,給藥后0.5h-6h內(nèi)P-PLK1表達明顯受抑制,24h P-PLK1表達恢復(fù)甚至高于對照組。3.PLK1抑制劑HS-10159能抑制NSCLC腫瘤細胞克隆形成能力,對A549細胞有明顯的放療增敏性,對LTEP,SPC細胞無明顯放療增敏性。PLK1抑制劑HS-10159對不同NSCLC細胞系有不同放療增敏性。4.三種NSCLC細胞隨單次給予的射線劑量的增加,P-PLK1蛋白表達表現(xiàn)為應(yīng)激性增高,給予單純藥物能抑制3種NSCLC P-PLK1的水平,給予藥物聯(lián)合放療能抑制單純放療所引起的P-PLK1水平的升高。5.PLK1抑制劑HS-10159聯(lián)合射線治療明顯增加NSCLC細胞有絲分裂G2/M期阻滯,較空白對照組細胞死亡及凋亡明顯增多,但PLK1抑制劑HS-10159聯(lián)合放療處理組較單純PLK1抑制劑HS-10159組細胞凋亡無明顯統(tǒng)計學(xué)差異。結(jié)論:1.臨床病例統(tǒng)計結(jié)果顯示PLK1蛋白的異常高表達與NSCLC的預(yù)后相關(guān),PLK1蛋白有可能作為預(yù)測NSCLC患者的預(yù)后指標(biāo)。2.PLK1抑制劑HS-10159抑制NSCLC細胞增殖能力及腫瘤細胞克隆形成能力,對不同NSCLC細胞系有不同放療增敏性。3.放療能導(dǎo)致NSCLC細胞P-PLK1蛋白表達短時間內(nèi)應(yīng)激性增高,PLK1抑制劑HS-10159聯(lián)合射線治療組能抑制單純射線所引起的P-PLK1蛋白表達的升高。PLK1抑制劑HS-10159聯(lián)合射線治療組明顯增加NSCLC細胞有絲分裂周期G2/M期阻滯,聯(lián)合治療組較單純藥物組細胞凋亡差異無明顯統(tǒng)計學(xué)意義。PLK1抑制劑HS-10159對NSCLC的放療增敏機制及參與的信號通路有待進一步探究。
[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.

【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R734.2

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1 李曉霞;PLK1抑制劑HS-10159在NSCLC中放療增敏機制的研究[D];天津醫(yī)科大學(xué);2016年

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本文編號：1869260