耐紫杉醇肺癌细胞BNX动物模型的建立  

Establishing a paclitaxel-resistant BNX mouse model of human lung cancer

作　　者：沙慧芳[1] 孙强玲[1] 杨晓华[1] 

机构地区：[1]上海市胸科医院胸部肿瘤研究所基础研究室,上海200030

出　　处：《第二军医大学学报》2011年第12期1296-1299,共4页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(81001037);上海市自然科学基金(10ZR1428100)~~

摘　　要：目的建立一种耐药性稳定的人肺癌BNX小鼠皮下移植瘤模型,为研究体内肿瘤耐药机制和逆转药物的筛选奠定实验基础。方法将体外培养的对数生长期的肺癌耐药细胞株A549-Taxol,接种在免疫缺陷小鼠的皮下,形成肿瘤,采用肿瘤组织块或肿瘤组织原代培养的细胞,分别于动物皮下接种致瘤;体内外交叉进行致瘤,提高耐药细胞株A549-Taxol的致瘤率和缩短致瘤时间。通过免疫组化鉴定该耐药细胞的特性。用MTT法检测细胞的耐药指数。结果耐药细胞接种于SCID小鼠4个月后在皮下形成肿瘤灶。用该肿瘤组织块或细胞悬液移植于SCID小鼠,通过反复3次后,细胞生长速度变快,2个月后皮下即可形成肿瘤。将该细胞移植于BNX小鼠,以同样的方法将瘤组织体内外交叉进行致瘤,最终BNX小鼠种植成功率达到80%。致瘤时间为3周。耐药细胞和组织块P-gp170、GST-π和MMP-7均高表达。A549-Taxol细胞的紫杉醇半数抑制浓度(IC50)是亲代A549细胞的520倍。结论初步建立了人肺腺癌耐药细胞(A549-Taxol)的动物模型,为肺癌临床个体化治疗及耐药逆转研究等提供了良好的实验动物平台。Objective To establish a paclitaxel-resistant BNX mouse model of human lung carcinoma,so as to provide evidences for studying chemoresistant mechanism and for screening of the reversal drugs in vivo.Methods The resistant model was produced by repeating a crossover subcutaneous injection of human lung cancer A549-Taxol cells and transplantation of tumor fragment into immune deficiency mice,so as to increase the tumor forming rate of A549-Taxol cells and shorten the tumor forming time.The expressions of GST-π,P-gp170 and MMP-7 were examined by immunohistochemical staining.The chemosensitivities of tumor cells to Taxol were tested and IC50 was measured by MTT.Results Tumor niduses were observed subcutaneously in SCID mice 4 months after injection of A549-Taxol cells,and then the tumor fragment or the tumor cells suspension were injected to SCID mice again.After 3 times of crossover injection,the tumor cells grew faster and tumor niduses were formed 2 months after injection.The same procedure was done in BNX mice.Finally,we achieved a successful rate of 80% in tumor implantation in BNX mice;the tumor niduses could be formed in 3 weeks.P-gp170,GST-π and MMP-7 expression was higher in the experiment group than that in the A549 control group.IC50 value of paclitaxel for A549-Taxol cells was 520 folds that of A549 cells.Conclusion We have successfully established paclitaxel-resistant lung carcinoma model in mice,which provides a new platform for further study on chemoresistant reversal strategy and individualized clinical treatment.

关 键 词：肿瘤抗药性 紫杉醇 动物模型 

分 类 号：R734.2[医药卫生—肿瘤]