耐紫杉醇肺原位小鼠移植瘤模型的建立和鉴定  

Establishment and characterization of a xenotransplant paclitaxel-resistant model of human lung cancer cells

作　　者：杨晓华[1] 郭莉莉 路丽明 YANG Xiaohua;GUO Lili;LU Liming(Central Laboratory,Shanghai Chest Hospital,Shanghai JiaoTong University,Shanghai 200030,China;Shanghai Institute of Immunology,Shanghai Jiaotong University School of Medicine,Shanghai 200025)

机构地区：[1]上海市胸科医院,上海交通大学附属胸科医院,实验中心,上海200030 [2]上海市免疫学研究所,上海交通大学医学院,上海200025

出　　处：《中国实验动物学报》2021年第3期275-283,共9页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金项目(81671579,82071856);上海市科学技术委员会实验动物研究项目(18140903800);上海交通大学转化医学协同创新中心合作资助项目(TM201522,TM201721);上海交通大学附属胸科医院协调创新项目(YJXT20190106);国家重点R&D项目重点专项(2017YFA0104600)。

摘　　要：目的建立肺原位移植耐紫杉醇肿瘤动物模型以及耐药机制的研究。方法利用本实验室已经获得的耐紫杉醇肺癌细胞株(A549-Taxol),测定其耐药指数和细胞对抗癌药物的敏感性后,采用穿刺法将约为每毫升5×10^(6)细胞注入小鼠肺部,3周后统计存活率以及成瘤率并且观察小鼠状态。RT-PCR和Western Blot技术检测肿瘤中耐药基因GST-π、P-gp170和MMP-7的mRNA和蛋白质的表达情况。结果A549-Taxol肺癌细胞的耐药指数为508倍;GST-π、P-gp170和MMP-7耐药蛋白表达明显升高(P<0.001),且A549-Taxol组肺癌细胞的侵袭能力显著高于A549组。用5×10^(6)细胞量进针5 mm的注射入肺部的存活率与成瘤率分别为100%和85%,成瘤后免疫组化显示耐药蛋白在A549-Taxol组裸鼠中明显表达,且耐药蛋白高于A549裸鼠组。结论GST-π、P-gp170和MMP-7的表达与肺癌紫杉醇耐药相关,初步建立了耐紫杉醇的肺癌原位动物模型,耐药细胞对紫杉醇的耐药性保持稳定并可用于后续的实验研究。Objective To establish xenotransplant paclitaxel-resistant models with A549 human lung cancer cells for the detection of drug resistance.Methods A549-taxol cells(5×10^(6)/mL)were injected into the lungs of nude mice.After 3 weeks,survival and tumor growth were recorded.MTT assays were performed to identify the drug-resistant index and drug susceptibility.Real-time PCR and western blotting were undertaken to detect Pgp170 and MMP-7 mRNA and protein expression,respectively.Results GST-π,P-gp170,and MMP-7 expression were significantly increased(P<0.001),and the invasiveness of GST-π-treated A549-taxol lung cancer cells was significantly higher than that of untreated A549-taxol cells and the A549-control group.The survival rate and tumor formation of A549-taxol cells were 100%and 85%,respectively.Immunohistochemistry showed that the drug resistance protein was significantly expressed in the A549-taxol group,and was higher than that in A549-control nude mice.Conclusions GST-π,P-gp170,and MMP-7 expression was associated with paclitaxel resistance in lung cancer.Lung orthotopic transplantation has been gradually established,and the current animal model of paclitaxel-resistant lung cancer remained stable,indicating its usefulness for future experiments.

关 键 词：肺癌 裸鼠 原位移植瘤模型 紫杉醇耐药 

分 类 号：Q95-33[生物学—动物学]