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目的探讨核糖体蛋白L39-L在肺癌A549细胞对阿霉素类药物耐药机制中的作用。方法以体外培养的阿霉素耐药和敏感人肺癌A549细胞株,Trizol法提取总RNA,Light Cycler RNA扩增法进行荧光定量RT-PCR,检测阿霉素耐药A549细胞株、敏感细胞株之间的RPL39-L基因表达差异。经对人肺癌A549敏感细胞株体外培养、稳定转染RPL39-L基因后,3H-Td R掺入法进行生存率检测分析核糖体蛋白L39-L转染A549细胞、载体转染A549细胞和空细胞株对阿霉素的耐药性差异。结果荧光定量RT-PCR结果显示,耐药性人肺癌A549细胞株比敏感的A549细胞核糖体蛋白L39-L转录水平高7.3倍。3H-Td R掺入测定生存率显示,与质粒载体转染细胞或空细胞相比,转染核糖体蛋白L39-L的细胞表现出增强的阿霉素耐药性。结论核糖体蛋白L39-L基因可能在肺癌A549细胞的耐药机制形成机制中有作用。
Objective To investigate the role of ribosomal protein L39-L in lung cancer A549 cells resistant to doxorubicin. Methods Adriamycin-resistant and susceptible A549 human lung cancer cell lines were cultured in vitro. The total RNA was extracted by Trizol method and the fluorescence quantitative RT-PCR was performed by Light Cycler RNA amplification. The expressions of adriamycin resistant A549 cell line, Differences in RPL39-L gene expression. The A549 cells were transfected with A549 cells and A549 cells were transfected with human lung cancer A549 cells in vitro and stably transfected with RPL39-L gene. 3H-TdR incorporation assay was used to determine the survival rate. Resistance of cell lines to doxorubicin. Results Fluorescent quantitative RT-PCR showed that the drug-resistant human lung cancer A549 cell line transcripts were 7.3-fold higher than the sensitive L49-L mRNA of A549 cells. 3H-TdR incorporation assay indicated that cells transfected with the ribosomal protein L39-L showed enhanced doxorubicin resistance compared to plasmid vector transfected or empty cells. Conclusion The ribosomal protein L39-L gene may play a role in the mechanism of drug resistance of lung cancer A549 cells.