Combination therapy in A549 cells.
Combination therapy in A549 cells.
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DOI：
10.1016/j.nucmedbio.2009.11.009
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发表时间：
2010-04
影响因子：
3.1
通讯作者：
Meng-hui Yuan;Jing Wang;Jinglan Deng;Zhe Wang;Wei-dong Yang;Guoquan Li;B. Ren
中科院分区：
文献类型：
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作者：
Meng-hui Yuan;Jing Wang;Jinglan Deng;Zhe Wang;Wei-dong Yang;Guoquan Li;B. Ren
背景与目的本实验研究了放射性核素{sup 131}I-RC-160与前体药物5-FC联合应用对共表达人生长抑素受体2基因（hSSTR 2）和大肠杆菌E.方法将hSSTR 2和CD基因克隆到双顺反子哺乳动物表达质粒中，并将其稳定转染A549细胞（pCIS-A549细胞）。抗生素筛选后，通过逆转录和聚合酶链反应（RT-PCR），Western blot，流式细胞术和免疫荧光分析确定稳定克隆中SSTR的表达。为了评估“工程化”A549细胞的体内靶向效率，将细胞皮下注射到裸鼠中，并在不同时间点评估{sup 99 m}Tc-RC-160的生物分布。通过肿瘤生长测量和免疫组化分析来评价{sup 131}I-RC-160和/或5-FC的肿瘤抑制作用。体内放射成像显示，与来自对照A549细胞的肿瘤相比，RC-160特异性靶向来自pCIS-A549细胞的肿瘤。结论hSSTR 2和CD基因在肿瘤细胞中的共表达可使肿瘤细胞对放射性同位素标记的RC 160和5 FC的超加和效应产生选择性增敏作用。这种方法为肺癌的治疗提供了一种潜在的治疗策略。
Background and aimWe investigated the anti-tumor effect induced by the combination of the radiotherapeutic agent {sup 131}I-RC-160 and the prodrug 5-FC in human non-small cell lung cancer (NSCLC) A549 cells that were co-expressing the human somatostatin receptor 2 gene (hSSTR2) and E. coli cytosine deaminase gene (CD).MethodsWe cloned both hSSTR2 and CD into a bicistronic mammalian expression plasmid and stably transfected it into A549 cells (pCIS-A549 cells). After antibiotic selection, SSTR expression in stable clones was determined by reverse transcription and polymerase chain reaction (RT-PCR), Western blot, flow cytometry and immunofluorescence analyses. To assess the in vivo targeting efficiency of the 'engineered' A549 cells, the cells were subcutaneously injected into nude mice and the biodistribution of {sup 99m}Tc-RC-160 was assessed at different time points. The tumor inhibitory effects of {sup 131}I-RC-160 and/or 5-FC were evaluated by measurement of tumor growth and immunohistochemical analysis.ResultsMultiple analyses demonstrated the successful expression of hSSTR2 in A549 cells. In vivo radioimaging revealed specific targeting of RC-160 to the tumors derived from pCIS-A549 cells when compared to those from control A549 cells. The tumor inhibitory rate of pCIS-A549 tumors in the {sup 131}I-RC-160 plus 5-FC-treated group was significantly higher than that in the single agent-treated group, control group and control tumors.ConclusionCo-expression of the hSSTR2 and CD genes in tumor cells can selectively sensitize these cells to the infra-additive effects of radioisotope-labeled RC-160 and 5-FC in vivo. This approach offers a potential therapeutic strategy for the treatment of lung cancer.