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[摘要] 目的 通过动物实验，观察S180瘤株对小鼠荷瘤形成的影响，探讨S180肉瘤细胞的致瘤机制。 方法 建立小鼠实体瘤模型，将昆明系小鼠36只，按体重随机分为3组，每组12只。空白组：每日灌胃给予蒸馏水，0.2 mL/10 g体重；模型组：皮下接种S180肉瘤细胞，制成小鼠实体瘤模型，其后每日口服灌胃1%羧甲基纤维素钠（CMC）溶液；环磷酰胺组：皮下接种S180肉瘤细胞，制成小鼠实体瘤模型后，注射0.02 g/（kgd）环磷酰胺注射液。观察各组肿瘤生长情况，计算平均重量；与模型组比较，环磷酰胺组计算肿瘤生长抑制率；采用蛋白免疫印迹法技术方法，检测抑癌基因P53、P21及凋亡抑制基因Bcl-2在S180肿瘤动物模型瘤体组织中的蛋白表达。 结果 模型组小鼠荷瘤成功，与空白组比较，模型组小鼠出现肿瘤，其瘤重为（1.5130.790）g，环磷酰胺组瘤重为（0.2480.253）g，两组比较差异有高度统计学意义（P 0.01），环磷酰胺抑瘤率为83.63%。在被检瘤体中，三种蛋白表达空白组与模型组、空白组与环磷酰胺组相比差异均有高度统计学意义（P 0.01）；与环磷酰胺组比较，模型组P53、P21抗体的蛋白表达量明显降低，差异有统计学意义（P 0.05），其中，P21表达差异有高度统计学意义（P 0.01），而Bcl-2变化不大，差异无统计学意义（P=0.66）。 结论 S180肉瘤细胞可能通过抑制P53与P21的表达而起到促进肿瘤形成的作用，该机制的研究可为筛选肿瘤治疗药物及肿瘤机制探讨等的肿瘤动物模型提供重要的实验依据。
[关键词] S180；蛋白免疫印迹法；P53；P21；Bcl-2
[中图分类号] R730 [文献标识码] A [文章编号] 1673-7210（2014）09（c）-0009-04
Effects and tumorigenic mechanism of S180 cell line in tumor-bearing mice model
CHI Shuping1 ZHANG Shilong1 CHEN Hongge1 LIU Jun1 DU Li1 SUN Jie1 JIANG ZhengJie2▲ CHENG Yun1▲
1.Department of Pharmacy Research Lab， 302 Hospital of PLA， Beijing 100039， China； 2.Health and Epidemic Prevention Team， Air Force Logistics Department of PLA， Beijing 100005， China
[Abstract] Objective To investigate the tumorigenic mechanism of sarcoma cell line （S180） through observing the effects of S180 on bearing tumor formation in mice. Methods Establishment of solid tumor model in mice， 60 Kunming mice were randomly divided into negative control group， model group and positive control group （Cyclophosphamide group） according to the body weight， 12 mice in each group. The negative control group was given distilled water， 0.2 mL/10 g. Except for the negative control group， subcutaneous injection with S180 sarcomas cells was done in the mice of other two groups. After the tumor model of mice was established， the model group mice were lavaged daily with 1% sodium carboxymethyl cellulose solution， and Cyclophosphamide group mice were injected Cyclophosphamide， 0.02 g/（kgd）. Then the tumor's volume and weight were measured， the tumor growth inhibition rate was calculated， and the expression of tissue protein of three kinds of antibody including tumor suppressor gene （P53 and P21） and apoptosis inhibiting gene （Bcl-2） were detected by Western blotting technology. Results The S180 cells planted tumor model of mice was established but not appearanced in negative control group. The tumor weight in model group and Cyclophosphamide group was （1.5130.790） g and （0.2480.253） g， respectively， it was significantly different （P 0.01） between the two groups. The inhibitory rate of positive control group was 83.63%. Compared with positive control group， the P53， P21 and Bcl-2 protein levels were significantly different in the model group and Cyclophosphamide group （P 0.01）. Compared with positive control group， the P53 and P21 protein level significantly decreased in model group （P 0.05）， the difference of P21 was highly statistically significant （P 0.01）， the changes of Bcl-2 were not statistically significant （P=0.66）. Conclusion The S180 sarcoma cells may promote tumor formation by inhibiting expression of P53 and P21. This mechanism can provide important experimental evidence for tumor animal model for screening drug and tumor mechanism.
[Key words] S180； Western blotting； P53； P21； Bcl-2
肿瘤是当前危害人类健康最主要的疾病之一，恶性肿瘤更是威胁生命的杀手。人类医学在对抗肿瘤的战争中，做了大量的科[(www. ) 专业提供论文代写和发表的服务,欢迎光临]学研究工作，其中最基本的就是肿瘤细胞模型与肿瘤动物模型的建立。Foley等[1]在1956年建立了可移植性小鼠S180肿瘤细胞株，将其注入CFW小鼠腹腔内保种传代。自细胞系建立以来，被广泛用于筛选肿瘤治疗药物及肿瘤机制探讨研究，同时荷S180小鼠也成为最常用的肿瘤动物模型之一[2]。由于S180瘤细胞株及其肿瘤动物模型在医学研究上具有重要的作用，因此，其在细胞内的作用机制亟待深入探究。本研究旨在通过建立肿瘤动物模型，观察肿瘤相关基因在肿瘤形成中的作用，以期在抗肿瘤动物模型研讨中减少和控制实验误差。
1 材料与方法
1.1 实验材料
1.1.1 药物与试剂 环磷酰胺购自山西普德药业有限公司（批号20070310）；BCA蛋白定量试剂盒购自Thermo（批号JL128100）；RPMI1640培养基、胎牛血清均购自Gibco公司，发光试剂盒购自Thermo公司，P53、P21、Bcl-2多克隆抗体、二抗均购自Santa公司。
1.1.2 动物与瘤株 昆明种健康小鼠，清洁级（2级），雌雄各半，体重（202）g，由军事医学科学院实验动物中心 |
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发表于 2018-8-18 21:47:18