【作者】 王宏；

【导师】 孙治君；

【作者基本信息】 重庆医科大学 ， 外科学， 2011， 硕士

【摘要】 第一部分乌司他丁和泰索帝对原代人乳腺癌细胞增殖、侵袭、凋亡以及PAFR、PDGFA、IGF-1R表达的影响目的:探讨乌司他丁(UTI)和泰索帝(TXT)对体外培养的原代人乳腺癌细胞的增殖、侵袭及凋亡的作用,以及对PAFR、PDGFA、IGF-1R基因扩增和蛋白表达水平的影响。方法:将培养的人原代乳腺癌细胞均衡分为4组:空白对照组,乌司他丁组,泰索帝组,乌司他丁+泰索帝组(其中UTI,800UI/ml; TXT,3.7μg/ml)。MTT法检测乳腺癌细胞的增殖能力;Transwell实验检测侵袭能力; RT-PCR和Western Blot分别检测各组细胞中PAFR、PDGFA、IGF-1R的基因扩增和蛋白表达水平;流式细胞仪检测各组细胞凋亡。结果:与对照和单独作用组相比,乌司他丁和泰索帝联合作用能够显著抑制原代培养的人乳腺癌细胞的增殖和侵袭,并且能诱导癌细胞凋亡。二者联合作用还能下调癌细胞中PAFR、PDGFA、IGF-1R的转录水平和蛋白的表达。结论:乌司他丁和泰索帝能够抑制乳腺癌细胞的增殖和侵袭,诱导细胞的凋亡,还能抑制细胞中PAFR、PDGFA、IGF-1R的表达,乌司他丁能够增强泰索帝的抗肿瘤作用。第二部分乌司他丁和泰索帝对人原代乳腺癌细胞裸鼠移植瘤生长及其PAFR、PDGFA、IGF-1R表达的影响目的:探讨UTI和TXT对原代人乳腺癌细胞裸鼠移植瘤的生长及PAFR、PDGFA、IGF-1R蛋白表达水平的影响。方法:利用原代人乳腺癌细胞建立裸鼠移植瘤模型,将模型裸鼠随机分为4组:Control组(生理盐水200μl/天/只),UTI组(1600UI/天/只),TXT组(20mg/7天/kg),UTI+TXT组(按单药剂量给药)。给药期间每隔3天测量肿瘤体积,并绘制生长曲线。用药3周后处死各组裸鼠,免疫组织化学检测各组PAFR、PDGFA、IGF-1R蛋白的表达。结果:裸鼠移植瘤生长曲线显示,对照组和UTI组肿瘤体积未见缩小,但UTI可延缓移植皮下瘤的增大。TXT及UTI+TXT组用药2周后肿瘤体积逐渐缩小,肿瘤抑制率为:TXT组88.44%,UTI+TXT组97.82%。与对照组相比,UTI、TXT能够明显下调移植瘤PAFR、PDGFA及IGF-1R蛋白的表达(p<0.05),且抑制作用UTI+TXT >TXT >UTI (p<0.05)。结论:UTI和TXT能够抑制裸鼠移植瘤体积的增大,UTI与TXT联合作用具有相加效应,能够下调裸鼠移植瘤中PAFR、PDGFA及IGF-1R蛋白的表达。更多还原

【Abstract】 PART 1 EFFECT OF ULINASTATIN AND TAXOTERE ON PROLIFERATION、INVASION、APOPTOSIS AND ON PAFR、PDGFA AND IGF-1R EXPRESSION OF PRIMARY HUMAN BREAST CANCER CELLObjective: To explore the synergistic effects of ulinastatin(UTI) and Taxotere(TXT) on proliferation、apoptosis and invasion of primary human breast cancer cell, as well as expression the PAFR, PDGFA, IGF-1R.Methods: The primary human breast cancer cell were divided into 4 groups: control group, UTI group, TXT group, UTI+TXT group (including UTI, 800UI/ml; TXT, 3.7μg/ml). The inhibitory effect of UTI and TXT on the cell proliferation and invasion of primary human breast cancer cell were detected by the MTT and Transwell assay, the effect of UTI and TXT on the cell apoptosis were detected by the Flow Cytometry (FCM). And then, the RNA and protein expressions of PAFR, PDGFA and IGF-1R were analyzed by Western Blot and RT-PCR.Results: The combination of ulinastatin and Taxotere synergistically decreased the cell proliferation and invasion of primary human breast cancer cell compared with control group and single medication group, as well as induced the cell apoptosis. Western Blot and RT-PCR results showed that the PAFR、PDGFA and IGF-1R expressions were significantly inhibited.Conclusions: UTI enhanced the inhibitory effect on proliferations of primary human breast cancer cell induced by TXT. Meanwhile, UTI showed synergistic effect with TXT by a potential mechanism of decreasing the expression of IGF-1R, PDGFA and PAFR. PART 2 EFFECT OF ULINASTATIN AND TAXOTERE ON TUMOR GROWTH AND EXPRESSION OF PAFR、PDGFA AND IGF-1R IN BREAST CANCER XENOGRAFT MODELSObjective: To investigate the effect of UTI and TXT on tumor growth and expression of PAFR, PDGFA and IGF-1R of human breast carcinomas in nude mice.Methods: Nude mice which injected with primary breast cancer cell in randomly were divided 4 groups: Control (Normal 200μl/day/mice), UTI (1600UI/day/mice), TXT (20mg/7days/kg), UTI (1600UI/day/mice) + TXT (20mg/7days/kg). Tumors Volume were measured and noted for growth curve every 3 days. Three weeks after treated, the all mice were sacrificed, the expression of PAFR, PDGFA and IGF-1R were measuring by immunohistochemistry.Results:UTI group could not decrease tumor volume, but can delay the increase of tumor volume in the tumor growth curves. Two weeks after grafting, TXT and UTI + TXT group gradually reduced tumor volume, inhibition rate: TXT 88.44%,UTI + TXT 97.82%. Compared with the control group, UTI and TXT can significantly down-regulate the protein expression of PAFR, PDGFA, IGF-1R (p <0.05), and the inhibition effect were UTI + TXT group> TXT group> UTI group (p <0.05). Conclusions: UTI and TXT could inhibit the growth of tumour and down-regulate the expression PAFR, PDGFA and IGF-1R of breast cancer xenografts tumors, UTI could enhance the synergistic effect of TXT.更多还原