膀胱癌5637细胞系的体外培养及荷瘤裸鼠实验动物模型的建立

发布时间：2017-12-31 08:06

本文关键词：膀胱癌5637细胞系的体外培养及荷瘤裸鼠实验动物模型的建立　出处：《北京协和医学院》2012年博士论文　论文类型：学位论文

【摘要】：背景：膀胱癌是泌尿系统最常见的肿瘤,临床上通常可分为局限于粘膜的浅表肿瘤和肌层浸润性肿瘤。治疗肌层浸润性膀胱癌的重要手段是全身化疗,但目前常用的化疗方案(GC、MVAC)疗效不佳,而且毒副作用比较严重。针对以上这些缺点,一种新的治疗方法开始被大家所重视,这就是分子靶向治疗。该方法具有选择性高、毒副作用小的特点。为了能够很好地评价靶向药物的疗效,最常用的方法就是建立膀胱癌的动物模型进行药物试验。目的：了解膀胱癌5637细胞系在体外的生长情况,并且建立动物模型以备进行药物试验。方法：取冻存的膀胱癌5637细胞进行复苏并传代培养,待细胞生长旺盛且形态较好时进行生长曲线的测定。将处于对数生长期的膀胱癌细胞制成细胞悬液,注射入2只裸鼠皮下,数周后取出肿瘤,将其切成小块移植入4只裸鼠皮下,观察肿瘤生长情况并测定肿瘤大小,取肿瘤组织进行HE染色后光镜下观察其结构。结果：膀胱癌5637细胞传代培养后生长速度良好。加入CCK-8试剂后,每日测定吸光度,以时间为横轴,吸光度为纵轴,绘制出生长曲线。结果显示,膀胱癌细胞d1-4处于对数生长期,增长速度快,活性佳,d4之后进入平台期。将膀胱癌细胞悬液注射入2只裸鼠皮下后约2-3周达到最大体积,之后将肿瘤组织块移植入4只裸鼠皮下,约2周肿瘤达到最大体积。结论：荷膀胱癌裸鼠异位移植瘤模型成瘤率高,生长迅速,肿瘤大小受实验操作影响较大。
[Abstract]:Background: bladder cancer is the most common tumor in urinary system, often clinically can be divided into the superficial tumors confined to the mucosa and muscle invasive tumor. Treatment of muscle invasive bladder cancer is an important means of systemic chemotherapy, but the common chemotherapy (GC, MVAC) poor efficacy and toxic side effects more serious. In view of the above shortcomings, a new method of treatment began to be valued, this is the molecular targeted therapy. This method has the advantages of high selectivity, small toxic side effect. In order to effectively evaluate the efficacy of drugs targeting the animal model, the most commonly used method is to establish the bladder cancer drug testing.
Objective: to understand the growth of the 5637 cell line of bladder cancer in vitro, and to establish an animal model for the drug test.
Methods: the recovery of cryopreserved and cultured from 5637 bladder cancer cells, when cell growth Washingce good morphology were determined. The growth curve of the logarithmic phase of growth of bladder cancer cells into cell suspension was injected into 2 nude mice, a few weeks after the tumor removed, cut it into pieces transplanted into 4 nude mice, growth of tumor was observed and measured tumor size, tumor tissues were observed with HE staining and its structure.
Results: the passage of bladder cancer 5637 cell culture growth speed is good. After adding CCK-8 reagent, the daily measurement of absorbance with time as the horizontal axis, the absorbance for the vertical axis, the growth curve was drawn. The results showed that bladder cancer D1-4 cells in logarithmic growth phase, fast growth, good activity, D4 after bladder cancer into the platform. The maximum volume of cell suspension was injected into the liquid reaches about 2-3 weeks after 2 nude mice, tumor tissue transplanted into 4 nude mice, tumor volume reached a maximum of about 2 weeks.
Conclusion: the ectopic tumor model of nude mice with bladder cancer has a high tumor rate and rapid growth, and the size of the tumor is greatly influenced by the experimental operation.

【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R737.14;R-332

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