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
1.Central Laboratory, Shanghai Chest Hospital, Shanghai JiaoTong University, Shanghai 200030, China. 2. Shanghai Institute of Immunology, Shanghai Jiaotong University School of Medicine, Shanghai 200025
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 × 106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 × 106 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 (5×106 / 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.
杨晓华,郭莉莉,路丽明.耐紫杉醇肺原位小鼠移植瘤模型的建立和鉴定[J].中国实验动物学报,2021,29(3):275~283.
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