【作者】 蒋丽琴；

【导师】 庄英帜； 曹建国；

【作者基本信息】 南华大学 ， 肿瘤学， 2008， 硕士

【摘要】 目的:观察过氧化物酶体增殖因子活化受体γ(Peroxisome Proliferator activated recettor gammar PPARγ)配体罗格列酮(Rosiglitazone ROZ)与顺铂(Cisplatin DDP)联合应用对人肺腺癌细胞系A549裸鼠移植瘤的作用,探讨ROZ活化PPARγ,下调NFκB、Bcl-2,从而诱导人肺腺癌细胞凋亡的作用机制。方法:体外培养人肺腺癌A549细胞,建立人肺腺癌(A549)细胞裸鼠移植瘤模型,28只荷瘤裸鼠随机分为7组, A组(生理盐水0.2ml),B组(DDP1mg/kg),C组(DDP4mg/kg),D组(ROZ10mg/kg),E组(ROZ30mg/kg),F组(ROZ10mg/kg +DDP1mg/kg),G组(ROZ 30mg/kg +DDP1mg/kg)。隔天腹腔注射给药,共8次,给药期间每4日测量皮下移植瘤的最长经和最短经,按标准公式:移植瘤体积(V)=最长径×最短径2×0.52计算瘤体积,绘制肿瘤生长曲线。实验末处死裸鼠,剥离皮下移植瘤,称量重量,计算瘤重抑制率;观察ROZ与DDP联合应用对人肺腺癌裸鼠移植瘤的治疗作用及化疗后人肺腺癌裸鼠移植瘤的病理学形态特点。移植瘤组织标本SP免疫组织化学检测PPARγ、NFκB、Bcl-2蛋白的表达。结果:人肺腺癌裸鼠移植瘤模型治疗实验结果显示:罗格列酮、顺铂及两药合用均能抑制裸鼠移植瘤的生长。DDP1mg/kg组及DDP4mg/kg组对移植瘤的瘤重抑制率分别为30.28%,65.85%,ROZ10mg/kg组及ROZ30mg/kg组对移植瘤的瘤重抑制率分别为38.38%,49.65%,呈剂量依赖性(P<0.01)。联合用药ROZ10mg/kg +DDP1mg/kg组及ROZ30mg/kg +DDP1mg/kg组抑瘤作用进一步增强,瘤重抑制率分别为52.11%,83.80%。ROZ30mg/kg与DDP1mg/kg联合应用具有协同效应(q=1.29)。免疫组化检测结果显示ROZ10mg/kg组及ROZ30mg/kg组与对照组比较, PPARγ蛋白表达上调,NFκB、Bcl-2蛋白表达下调,差异具有显著性(P<0.05)。联合用药组该调节作用进一步增强(P<0.001)。结论:1)罗格列酮能抑制人肺腺癌裸鼠移植瘤的生长。2)罗格列酮能增强顺铂对人肺腺癌裸鼠移植瘤的生长抑制作用,呈剂量依赖性。3)罗格列酮增强顺铂抑制人肺癌裸鼠移植瘤生长作用可能与其上调PPARγ,下调NFκB、Bcl-2蛋白表达相关。更多还原

【Abstract】 Objective:To observe the synergistic effects of Rosiglitazone and Cisplatin on the growth of lung adenocarcinoma A549 xenogragft in nude mice, and to investigate the mechanism of the therapeutic and synergistic action whether is involved in activating PPARγ.Methods:Human lung carcinomal cell line (A549) cells were cultured in vitro, then A549 cells were transplanted in nude mice. Twenty-eight xenogragfts in nude mice were established successively. They were then randomly divided into seven groups. A(N.S group),B(Cisplatin 1mg/kg),C(Cisplatin 4mg/kg),D(Rosiglitazone 10mg/kg), E(Rosiglitazon30mg/kg),F(Rosiglitazon10mg/kg+Cisplatin1mg/kg),G(Rosiglitazone30mg/kg+Cisplatin1mg/kg).Seven days after implantation, different treatments were served. The volumes of the xenogragfts were measured respectively during the therapy time. The tumor growth inhibiting rates of each group were calculated. Moreover, the curves of tumor growth were drew. Xenogragft was subjected to histological examination. Subcutaneous xenogragft PPARγ, Bcl-2 and NFκB were determined by immunohistochemistry method.Results: Therapeutic trial of human lung carcinoma xenografts in nude mouse model indicated that In each treatment group, tumor growth was suppressed significantly, intraperitoneal injection of Cisplatin, Rosiglitazon and their combination resulted in a significant inhibition on the growth of human lung carcinoma xenografts A549 in vivo(P<0.01), The tumor weight inhibitory rate of DDP at 1mg/kg, 4mg/kg was 30.28%, 65.85%,respectively. The tumor weight inhibitory rate of ROZ at 10mg/kg, 30mg/kg was 38.38%, 49.65% respectively. The tumor weight inhibitory rate of combination groups was 52.11%, 83.80%. The effect of inhibiting tumor growth was significantly strengthened. The combination group (Rosiglitazone 30mg/kg+ Cisplatin 1mg/kg) showed synergia effect on inhibiting the growth of the xenografts with a q value of 1.29. Campare to control group, Bcl-2 and NFκB expression of subcutaneous xenografts was obviously down-regulated and the PPARγwas up-regulated by Rosiglitazone (P<0.05) and the combined group(P<0.001).Conclusion:1) Rosiglitazone can inhibit the growth of human lung carcinoma xenograft in nude mice.2) Rosiglitazone combined with Cisplatin can significantly inhibit the growth of human lung carcinoma xenograft in nude mice in dose-dependent manner.3) The inhibition of growth by Rosiglitazone combined with Cisplatin are associated with up-regulation of PPARγprotein level expression and down-regulation of NFκB、Bcl-2protein level expression.更多还原