小鼠原位移植性肝癌模型的建立及生物学特性

Establishment of orthotopic transplantation tumor model of liver cancer and its biological features

目的:探讨建立小鼠原位移植性肝癌模型的可行性并评价其生物学特性。方法:沿腹中线开腹后将5×106个H22细胞接种到小鼠(17只)肝脏实质内,分别于接种后第13天(n=8)和小鼠死亡时(n=9)取主要脏器进行形态学观察和细胞增殖核抗原(PCNA)及免疫Ⅷ因子相关抗原(FⅧRag)免疫组织化学染色。结果:小鼠平均生存期为18.8 d。接种后第13天,肝内肿瘤成瘤率为100%(8/8),肺转移率为37.5%(3/8),50%(4/8)的小鼠产生腹水,腹水量为(6.25±3.67)mL,肿瘤体积为(0.35±0.14)cm3。自然死亡时,肺转移率为100%(9/9),100%(9/9)小鼠产生腹水,腹水量为(10.88±1.92)mL,明显多于接种后第13天时的腹水量(P<0.01);肿瘤体积为(0.61±0.31)cm3,明显大于接种后第13天时的肿瘤体积(P<0.05)。小鼠死亡时大多数瘤组织呈单结节状,异型性明显,肿瘤增殖指数和微血管密度分别为(71.39±20.74)%和(39.25±13.71)/高倍视野。结论:原位移植性肝癌模型制备成功率高,肿瘤肺转移率高,肿瘤组织具有丰富的微血管和旺盛的增殖能力。

Objective To explore the feasibility of orthotopic transplantion tumor model of hepatoma in mice and evaluate the biological features. Methods 5 × 10^6 H22 cells were inoculated into the liver of mice after anesthetizing. The liver, spleen, lung and kidney were taken from the mice 13 d after inoculation （n=8） or from the died mice （n=9） for morphological evaluation. Proliferating cell nuclear antigen （PCNA） and factor Ⅶ related antigen （FⅦRag） were analyzed with immunohistoehemical staining. Results The average survival time of the mice was 18. 8 d. Until the 13th day after inoculation, the successful rate of orthotopic transplantation tumor （model of liver cancer） was 100~ （8/8）, spontaneous lung metastatic rate was 37.5% （3/8）. Fifty percent of mice （4/8） produced ascites, and the average amount of ascites was （6. 25±3.67） mL. The average size of the tumor was （0.35 ±0.14） cm^3. In the natural died mice, spontaneous lung metastatic rate was 100% （9/9）, 100% of mice （9/9） produced ascites. The average volume of aeites was （10.88±1.92） mL, which was evidently more than that of the 13th day （P〈0.01）. The average size of the tumor was （0.61±0.31） cma, which was significantly bigger than that of the 13th day （P〈0.05）. When the mice died, most of tumor nodules were single and the cellular atypia was apparent. The proliferation index and microvessel density of tumor tissue were 71.39%± 20. 74% and （39.25±13.71） /400 high magnification respectively. Conclusion The suecessful rate of orthotopic transplantation tumor model of liver cancer is high. The tumor possess high lung metastatic rate and plenty of microvessels and strong proliferative ability.