热化疗联合对人肺腺癌耐药细胞株A549/DDP作用及其机制的实验研究

目的观察热化疗联合对人肺腺癌耐药细胞株A549/DDP的影响及其机制,为临床上应用热化疗联合治疗非小细胞肺癌提供理论依据。方法CCK-8法检测A549、不同温度下A549/DDP细胞对顺铂的敏感性;流式细胞仪分析热疗对A549/DDP细胞周期分布的影响;RT-PCR检测热疗对A549/DDP细胞ERCC1表达的影响。结果亲代人肺腺癌细胞株A549对顺铂的敏感性明显高于耐药细胞株A549/DDP,不同温度加热后A549/DDP细胞对顺铂的敏感性均有提高;热疗尤其是热化疗后G2/M期细胞明显增多,G0/G1期细胞明显减少(P<0.01);热疗后A549/DDP细胞ERCC1mRNA表达未见明显改变。结论热疗能提高人肺腺癌耐药细胞株A549/DDP对顺铂的敏感性,热化疗具有协同作用。热化疗协同作用机制可能主要是诱导细胞周期阻滞,抑制细胞增殖,和DNA修复基因ERCC无直接相关性。

Objective To observe the effects and mechanisms of hyperthermia therapy combined with chemotherapy on multidrug- resistant human lung edenocarcinoma ceil line A549/DDP, it will provide theory evidence in thennochemotherapy for clinic. Methods A549 or A549/DDP treated with different temperature were detected of sensitivity to cisplatin with CCK- 8 assay. After hyperthennia, A549/DDP cell cycle was analyzed with flow cytometry and ERCClmRNA expression was detected using RT-PCR. Results Parent human lung adenecarcinoma ceil A549 was more sensitive to cisplatin than drug- resistant ceil line A549/DDP. Treated with different temperature, drug sensitivity of A549/DDP to cisplatin all increased. Hyperthermia had significant effect on cell cycle with increase of G2/M phase population and reduction of GO/G1 phase population, and thennochemotherapy was more notable（ P 〈 0.01 ）. After hyperthennia, the expression of ERCC1 mRNA did not change. Conclusion Hyperthermia can significantly enhance the sensitivity to cisplatin in human muhidrug- resistant lung cancer cell line A549/DDP. Tharmochemotherapy has synergistic effects of inhibiting ceil proliferatian. The major synergistic mechanisms of thermochemo.herapy are that they can induce cell cycle arrest,inhibit cell proliferation , but have no correlation with gene ERCC1.