重组人血管内皮抑素（Endostar）增加放疗敏感性的机制及临床应用探讨

发布时间：2018-02-02 04:45

本文关键词： 重组人血管内皮抑素肺癌细胞周期凋亡血管新生肺癌放射治疗 Endostar AQP-1 HIF-1α MVD 非小细胞肺癌恩度/血管内皮抑素同步放化疗不良反应　出处：《武汉大学》2014年博士论文　论文类型：学位论文

【摘要】：第一部分Endostar联合放疗对人肺腺癌A549细胞作用及其机制的研究目的：研究Endostar(ES)对人肺腺癌A549细胞增殖、细胞周期、凋亡等生物学行为的影响及其可能机制。方法：常规培养细胞,实验分为对照组、ES组、放疗(RT)组、ES联合RT组。MTT方法测定对照组及实验组细胞增殖抑制率；流式细胞仪分析细胞周期变化；Hoechst染色法观察细胞凋亡率；ELISA及RT-PCR法检测HIF-1α、VEGF、bFGF、 TGF-β1蛋白表达。结果：与对照组相比,ES抑制A549细胞增殖,且呈剂量依赖性,在25μg/L时达到76.7%；ES可显著阻滞A549细胞至G2/M期和S期(P0.05)；联合组细胞凋亡率明显高于其他各组(P0.05)；联合组HIF-1α、VEGF、bFGF、TGF-β1相对表达量明显低于其他各组(P0.05),呈剂量依赖性,且其抑制作用随时间延长而增强。结论：ES具有抑制肺癌增殖、阻滞周期的作用,与放疗联用时可通过抑制HIF-1α、VEGF、bFGF、TGF-β1从而增强放疗的促肿瘤凋亡效应。第二部分Endostar联合放疗对lewis肺癌小鼠移植瘤的作用及其机制的研究目的：探讨ES联合放疗对移植瘤生长及血管新生的影响。方法：实验分组：对照组、ES组、RT组及联合组。绘制肿瘤生长曲线,计算抑瘤率；免疫组化(SP法)测定VEGF水平及微血管密度(microvessel density,MVD); RT-PCR检测AQP-1、HIF-1αmRNA表达。结果：与对照组相比,ES组、RT组及联合组移植瘤生长速度、瘤体质量、MVD显著降低(P0.05),其中联合组较其他各组降低最为明显(P0.05)；与对照组相比,ES组AQP-1、HIF-1α水平降低,但无统计学意义(P0.05),联合组AQP-1表达显著增高、HIF-1α水平显著降低,与其他各组相比,差异显著(P0.05)。结论：Endostar联合放疗能明显抑制lewis肺癌小鼠移植瘤的生长,其机制可能是下调HIF-1α、VEGF的表达、上调AQP-1的表达,抑制血管新生。第三部分恩度联合同步放化疗治疗局部晚期非小细胞肺癌的临床观察目的：观察恩度(重组人血管内皮抑素,Recombinant human endostatin, Endostar, YH-16)联合同步化疗、放疗方案治疗局部晚期非小细胞肺癌(non-small cell lung cancer, NSCLC)的近期临床疗效和安全性。方法：回顾性分析了59例经组织病理学确诊的局部晚期非小细胞肺癌患者的临床资料。依照纳入标准,分为恩度联合同步放化疗组(试验组)30例和同步放化疗组(对照组)29例,进行相关数据对比分析,观察以客观有效率(RR)、临床受益率(CBR)、1年生存率和1年无进展生存率为近期疗效评价指标。评价血液学毒性、消化道反应、心脏毒性、放射性肺炎和放射性食管炎等毒副反应。结果：试验组与对照组的客观有效率(CR+PR)分别为66.7%和37.9%,差异有统计学意义(P0.05),临床受益率(CBR)分别为90%和69%,差异有统计学意义(P0.05),1年生存率分别为70.0%、53.3%,1年无进展生存率58.6%、44.8%,差异无统计学意义(P0.05)。主要的不良反应有骨髓抑制、恶心呕吐、急性放射性肺炎及食管炎、心脏毒性,两组不良反应差异均无统计学意义。结论：同步放化疗联合恩度治疗晚期非小细胞肺癌的临床治疗近期疗效好,毒副反应可接受。
[Abstract]:Effect of first part Endostar combined with radiotherapy on human lung adenocarcinoma A549 cells and its mechanism Objective : To study the effects of Endostar ( ES ) on the proliferation , cell cycle and apoptosis of human lung adenocarcinoma A549 cells and its possible mechanism . Methods : Normal cultured cells were divided into control group , ES group , radiotherapy ( RT ) group and ES combined RT group . The proliferation inhibition rate of the control group and the experimental group were determined by MTT method . The apoptosis rate was observed by flow cytometry . The expression of HIF - 1伪 , VEGF , bFGF and TGF - 尾1 was detected by ELISA and RT - PCR . Results : Compared with the control group , ES inhibited the proliferation of A549 cells and reached 76.7 % at 25 渭g / L ; ES significantly blocked A549 cells to G2 / M phase and S phase ( P0.05 ) . The relative expression of HIF - 1伪 , VEGF , bFGF and TGF - 尾1 in the combined group was significantly lower than that in other groups ( P0.05 ) . Conclusion : ES has the function of inhibiting the proliferation and block cycle of lung cancer , and can enhance the tumor - promoting effect of radiotherapy by inhibiting HIF - 1伪 , VEGF , bFGF and TGF - 尾1 when combined with radiotherapy . Effect of the second part of Endostar combined with radiotherapy on the transplanted tumor of lewis lung cancer mice and its mechanism Objective : To investigate the effects of combined radiotherapy and radiotherapy on tumor growth and angiogenesis . Methods : Experimental group : control group , ES group , RT group and combined group . Tumor growth curve was drawn , tumor inhibition rate was calculated and VEGF level and microvessel density ( MVD ) were measured by immunohistochemistry ( SP method ) ; AQP - 1 and HIF - 1伪 mRNA expression were detected by RT - PCR . Results : Compared with the control group , the growth rate , tumor mass and MVD were significantly lower in the ES group , the RT group and the combined group ( P0.05 ) . Compared with the control group , the expression of AQP - 1 and HIF - 1伪 in the ES group was significantly higher than that in the control group ( P0.05 ) . Conclusion : Endostar combined with radiotherapy can significantly inhibit the growth of lewis lung cancer mouse transplanted tumor . The mechanism may be to downregulate the expression of HIF - 1伪 and VEGF , up - regulate the expression of AQP - 1 and inhibit angiogenesis . The Clinical Observation of the Third Part Ndegree Combined with Concurrent Chemoradiotherapy in the Treatment of Locally Advanced Non - small Cell Lung Cancer Objective : To observe the recent clinical efficacy and safety of the combination of chemotherapy and radiotherapy in the treatment of locally advanced non - small cell lung cancer ( NSCLC ) . Methods : The clinical data of 59 patients with locally advanced non - small cell lung cancer diagnosed by histopathological examination were analyzed retrospectively . The results showed that the objective response rate ( RR ) , clinical benefit rate ( CBR ) , 1 - year survival rate and 1 - year progression - free survival rate of the patients were 66.7 % and 37.9 % , respectively . The results showed that the objective response rate ( CR + PR ) of the test group and the control group were 66.7 % and 37.9 % , respectively . The difference was statistically significant ( P0.05 ) . The main adverse reactions were bone marrow suppression , nausea and vomiting , acute radiation pneumonia , esophagitis and cardiotoxicity . Conclusion : The combination of synchronous radiotherapy and chemotherapy in the treatment of advanced non - small cell lung cancer has a good effect in the near term , and the toxic side effect is acceptable .

【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R730.55

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