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目的:建立小鼠肝癌休眠模型并验证休眠细胞的存在。方法:30只昆明种小鼠均右侧腋下接种1×106个H22腹水型肝癌细胞,常规喂养2周,第15天选择表皮肿瘤直径>0.5cm的小鼠,进行姑息性手术,将肉眼可见肿瘤组织全部切除。手术后的小鼠再常规喂养8周,如接种处未见肿瘤组织生长,即认为肝癌小鼠休眠模型建立成功(按照平均寿命折算,小鼠生存期8周,即相当于人类寿命5年)。其后模型小鼠分为复发对照组和复发实验组,复发对照组小鼠常规喂养;复发实验组模型小鼠给予连续外伤刺激。6周后处死全部模型小鼠,接种部位取材,组织固定和HE染色观察肿瘤再生成情况确定休眠细胞的存在。结果:肝癌细胞接种于小鼠并行手术后,有20%小鼠成瘤,80%处于休眠状态。复发实验组给予连续外伤刺激6周后,100%小鼠成瘤,而复发对照组只有8.3%,两组差异有统计学意义,P=0.000。结论:本方法成功建立了小鼠肝癌休眠模型,并用外伤刺激引起肝癌休眠细胞增殖,验证了休眠肿瘤细胞在体内的存在。
Objective: To establish a mouse model of liver cancer dormancy and verify the presence of dormant cells. Methods: Thirty Kunming mice were inoculated with 1 × 106 H22 ascites hepatoma cells on the right side of the axilla. The mice were routinely fed for 2 weeks. On the 15th day, mice with epidermal tumors> 0.5 cm in diameter were selected for palliative surgery. Tumor tissue can be seen all removed. After surgery, the mice were routinely fed for another 8 weeks. If there was no tumor growth at the inoculation site, the mice model of liver cancer was considered to be successful (based on the average life expectancy, the survival of mice was 8 weeks, which is equivalent to 5 years of human life) . Subsequent model mice were divided into recurrent control group and relapse experimental group, and relapsed control group mice were fed routinely. The mice in relapse experimental group were given continuous traumatic stimulation. After 6 weeks, all the mice were sacrificed, the inoculation site was taken out, tissue fixed and HE staining was used to observe the tumor regeneration to determine the presence of dormant cells. RESULTS: Twenty percent of the mice were tumorigenic and 80% were in a dormant state after the hepatoma cells were inoculated into the mice in parallel. After 6 weeks of continuous trauma stimulation, 100% of the mice in the recurrent experimental group developed tumor, while the recurrent control group only had 8.3%. There was a significant difference between the two groups (P = 0.000). Conclusion: This method successfully established mouse liver cancer dormancy model and stimulated with traumatic stimulation of the proliferation of dormant cells of liver cancer to verify the existence of dormant tumor cells in vivo.