A549肺腺癌多细胞球体药敏实验、Mdr1、MRP表达以及^(99m)Tc-MIBI摄取研究  

Research of Chemosensitivity, Mdr1、MRP Expression and ^(99m)Tc-MIBI Uptake in A549 Multicellular Spheroids

作　　者：史德刚[1] 黄钢[2] 苗积生[3] 林祥通[1] 

机构地区：[1]复旦大学附属华山医院核医学科 [2]上海第二医科大学附属仁济医院核医学科,上海200001 [3]上海市肺科医院核医学科,上海200433

出　　处：《复旦学报（医学版）》2005年第4期463-466,共4页Fudan University Journal of Medical Sciences

摘　　要：目的探讨A549肺腺癌多细胞球体的多药耐药基因(mdr1)和多药耐药相关蛋白(MRP)基因表达,甲氧基异丁基异腈(MIBI)摄取结果能否预测化疗效果。方法①血浆高峰浓度的化疗药物顺铂(DDP)、长春花碱酰胺(VDS)、氟尿嘧啶(5-氟尿嘧啶,5-FU)、羟喜树碱(HCP)、丝裂霉素(MMC)、多柔比星(阿霉素,ADM)分别进行药物敏感性实验。②单层贴壁细胞培养。96孔板1个,每组4孔,2×104细胞/孔。③MTT检测观察疗效。细胞加化疗药后培养48h,然后加MTT5mg/mL,每孔20μL,4h后用酶标仪(490nm)测量吸光度值。④RT-PCR检测A549细胞和MCF-7耐药株的mdr1和MRP表达,β2-MG作内参照。⑤96孔板内每孔一个球体,每孔掺入99mTc-MIBI9.25×104Bq(2.5μCi),分别于60和120min结束掺入并测量γ计数。结果①A549细胞药敏实验对DDP不敏感,对MMC、VDS、ADM、5-FU、HCP敏感。②MCF-7长春新碱耐药株(MCF-7/VCR)对DDP、VDS不敏感,对MMC、ADM、5-FU和HCP较敏感。③A549细胞及球体的Mdr1/β2-MG为0,MRP/β2-MG分别为0.76和0.62;MCF-7耐药细胞的mdr1/β2-MG和MRP/β2-MG分别为35和4.36。④A549多细胞球体对99mTc-MIBI120min的摄取值高于60min摄取值(P<0.05)。结论A549多细胞球体的mdr1和MRP表达水平较低以及MIBI摄取阳性提示无多药耐药性产生,与化疗药物敏感性检测结果基本一致。A549细胞对DDP耐药说明除mdr1和MRP之外,还存在与转运蛋白间接相关的其他耐药机制。Purpose To investigate whether the gene expressions of multidrug resistance gene (mdr1) multidrug resistance-associated protein (MRP) and uptake of methoxyisobutylisonitrile (MIBI) in A549 multicellular spheroids (MTS) could predict chemotherapeutic effect. Methods ① Chemosensitivity test was done individually with peak plasma concentration of cisplatin (DDP)?vindesine (VDS)?5-fluorouracil (5-FU)?hydroxy camptothecin (HCP)?mitomycin C (MMC) and doxorubicin.② Cell culture in single layer was done in 96 hole plate,every group was consist of 4 holes each contained 2×104 cells.③ MTT assay was used to evaluate the treatment effect.Four hours after mixed the chemotherapeutic drugs into holes for 48 h and added 20 μL of MTT with 5 mg/mL,absorbency was measured in 490 nm.④ Expression of mdr1 and MRP in A549 cells and MCF-7/vincristine (VCR)resistance cells were tested by RT-PCR method,with β2-MG acted as internal conference.⑤ Every MTS was placed in every hole of 96 hole plate and mixed with 9.25×104 Bq(2.5 μCi) of 99mTc-MIBI for 60 and 120 min individually,then γ counts were measured. Results ① A549 cells were insensitive to DDP,but sensitive to MMC?VDS?ADM?5-FU and HCP.② MCF-7/VCR cells were insensitive to DDP and VDS,but minor sensitive to MMC,ADM,5-FU and HCP.③ The ratios of mdr1/β2-MG in A549 cells and MTS were 0,and those of MRP/β2-MG were 0.76 and 0.62 individually.The ratios of mdr1/β2-MG and MRP/β2-MG in MCF-7/VCR cells were 35 and 4.36 individually.④ The uptakes of 99mTc-MIBI in A549 MTS for 120 min were more than those for 60 min (P<0.05). Conclusions The lower gene expressions of mdr1 and MRP in A549 MTS and positive uptake result of 99mTc-MIBI reflected that the cells didn's have multidrug resistance,which was consistent with its chemosensitivity test.For the A549 cells resisting to DDP,apart from mdr1 and MRP,other resistance mechanisms which had indirected relation to pump protein may be existed.

关 键 词：^99mTc-MIBI 多细胞球体 药敏实验 Mdr1 MRP 肺腺癌 摄取 A549细胞 β2-MG 多药耐药相关蛋白 甲氧基异丁基异腈 RT-PCR检测 MCF-7 药物敏感性实验 化疗药物敏感性 mdr1 多药耐药基因 血浆高峰浓度 长春花碱酰胺 5-氟尿嘧啶 

分 类 号：R541.4[医药卫生—心血管疾病] R737.31[医药卫生—内科学]