不同方法建立的人大细胞肺癌裸鼠移植瘤模型的生物学特点

Biological Characteristics of Three Nude Mouse Models of Human Large Cell Lung Cancer Established by Different Methods

目的比较三种常用的皮下移植瘤造模方法建立的人大细胞肺癌NCI-H460裸鼠移植瘤模型的不同生物学特点,为不同的研究寻找合适的造模方法提供实验依据。方法分别用NCI-H460细胞,NCI-H460移植瘤组织块和移植瘤匀浆液对于BALB/c-nu/nu裸鼠建立皮下移植瘤模型,运用一般生物学指标观察三种移植瘤的成瘤率、瘤重、倍增时间和组织形态;采用全自动生化分析仪检测其外周血中丙氨酸氨基转氨酶(ALT)、天冬氨酸氨基转移酶(AST)、血糖(G1u)、尿素氮(BUN)和肌酐(CREA)等生化指标;利用血球分析仪检测白细胞总数(WBC)并进行分类,最后体外对其腹腔巨噬细胞吞噬活性和NK细胞活性进行了考察。结果本实验中细胞法和匀浆法的成瘤率及肿瘤生长速率显著高于埋块法且其生长更为均一,差异较小。接种5周后,与正常裸鼠比较,三组荷瘤小鼠血液中ALT、AST显著升高,BUN、CREA显著降低,埋块组的AST和BUN两项指标显著高于其他两荷瘤组。此外,接种2周后,荷瘤裸鼠的GLU显著低于正常裸鼠,匀浆液组的GLU降得最低。白细胞中,三种方法组荷瘤小鼠血液中LYM%、MN%、HGB均有降低,匀浆液组和细胞培养组的荷瘤小鼠血液中WBC、NEUT%、PLT显著高于埋块组。免疫细胞活性方面,两种细胞均呈现出正常细胞组>匀浆组>细胞组>埋块组的趋势。结论细胞培养法接种数量可控,肿瘤生长均匀,适合建立不同实验需求的移植瘤模型,组织块移植法适于建立中药抗肿瘤筛选的动物模型,而匀浆液移植法则不推荐使用。裸鼠的生理生化状态和免疫功能与肿瘤的生长有密切的关系。

Objective To compare the biological characteristics of three modeling methods to establish human large cell lung cancer（LCLC） NCI-H460 xenograft models in nude mice,and provide an experimental basis for finding a suitable modeling approach for different studies.Methods Cultured NCI-H460 cells,NCI-H460 tumor tissue pieces and tumor homogenate were used respectively to establish subcutaneous tumor models in BALB/c-nu/nu nude mice.The tumor formation rate,tumor weight,doubling time and morphology were observed.Blood alanine aminotransferase（ALT）,aspartate aminotransferase（AST）,glucose（G1u）,urea nitrogen（BUN） and creatinine（CREA） were analyzed using an automatic biochemical analyzer.WBC count and classification were determined,and finally the phagocytic activity of macrophages and NK cell activity in vitro were assessed.Results The tumor formation rate,tumor growth rate and uniformity in the cultured cells and tumor homogenate groups were significantly better than those of the tissue transplantation group.Five weeks after inoculation,the blood ALT and AST were significantly increased,while BUN,CREA were significantly lower in all the three groups of tumor-bearing mice,the AST and BUN of the tissue transplantation group were significantly higher than those of the other two groups.However,the Glu of tumor-bearing mice decreased significantly 2 weeks after inoculation.The LYM%,MN% and HGB of the peripheral blood of the three groups were lower than those in the normal control group,and WBC,NEUT%,PLT of the cultured cell and homogenate groups were significantly higher than those of the tissue transplantation group.The immune cell activity of the macrophages and NK cells showed a lowering trend in an order of normal control grouphomogenate groupcultured cell grouptissue transplantation group.Conclusions The modeling method with cultured cells is controllable in number and uniform of tumor growth.It is suitable for the establishment of different experimental xenograft models.The tissue transplantation can meet the need for the establishment of animal models to serve Chinese anti-cancer drug screening,and the homogenate inoculation method is not recommended.The physiological and immunological characteristics of the mouse models have a close relationship with tumor growth.