【作者】 吴朔；

【导师】 李宁；

【作者基本信息】 遵义医学院 ， 内科学， 2010， 硕士

【摘要】 目的：1、研究JS-K对耐顺铂人肺腺癌细胞A549／DDP耐药的逆转作用。2、研究JS-K对人肺腺癌细胞A549及耐顺铂人肺腺癌细胞A549／DDP多药耐药相关基因GST-πmRNA表达的影响。方法：1、采用MTT法检测JS-K联合顺铂对耐顺铂人肺腺癌细胞A549/DDP的抑制作用及逆转倍数。2、采用real-time RT-PCI法检测不同浓度JS-K处理前后人肺腺癌细胞A549及耐顺铂人肺腺癌细胞A549／DDP的GST-πmRNA的表达。结果：1、顺铂对人肺腺癌细胞A549及耐顺铂人肺腺癌细胞A549／DDP(48h)的IC50值分别为1.702±0.100μg/ml、9.943±0.063μg/ml,其抑制作用差异有统计学意义(P<0.05)；JS-K对人肺腺癌细胞A549及耐顺铂人肺腺癌细胞A549／DDP均有抑制作用,其抑制作用差异无统计学意义(P>0.05)。JS-K对耐顺铂人肺腺癌细胞A549／DDP的IC10值为1.042±0.124μM。以JS-K IC10值以下的浓度为无毒剂量,选择0.1μM、0.3μM、1μM三种不同浓度的JS-K联合顺铂可以增加耐顺铂人肺腺癌细胞A549／DDP对顺铂的敏感性,使顺铂的IC50值分别降至1.963±0.238μg/ml.1.371±0.119μg/ml、1.083±0.055μg/ml,差异有统计学意义(P<0.05)。对顺铂的耐药逆转倍数分别为5.066、7.254、9.178。2.0.1μgM、0.3μM、1μM三种不同浓度的JS-K处理两种细胞48h后检测其耐药基因GST-πmRNA表达,其GST-πmRNA的相对表达量均较对照组明显下降,且呈浓度依赖性,差异有统计学意义(P<0.05)。结论：JS-K可以增强耐顺铂人肺腺癌细胞A549/DDP对顺铂的敏感性,逆转其耐药。这可能与下调多药耐药相关基因GST-πmRNA表达有关。更多还原

【Abstract】 Objective:To investigate the reversal effects of the nitric oxide (NO) prodrug JS-K on GST-induced cisplatin resistant cell line A549/DDP. To investigate the impact of JS-K to the expression of GST-πmRNA on A549/DDP and A549 cells. Methods:A549 and A549 /DDP cells were cultured with DDP alone or in combination with JS-K respectively. The inhibition of DDP and the inhibition of DDP in combination with JS-K were assessed with MTT and the IC50 values were calculated. The expression of GST-πmRNA was detected by real-time quantitative reverse transcription-polymerase chain reaction (real-time RT-PCR). Results:The IC 50 value was 1.702±0.100μg/ml,9.943±0.063μg/ml when A549 and A549/DDP cells treated with DDP. The difference between the two groups had no statistical significance (P<0.05). JS-K also has an inhibition effect on both A549 and A549/DDP cells. The difference between the two groups had no statistical significance (P> 0.05).The JS-K IC 10 value was 1.042±0.124μM. Chose JS-K IC10 value as its non-toxic dose. DDP combined with JS-K (0.1μM,0.3μM, 1μM) that bellowed its IC 10 value could significant decrease the IC50 value to 1.963±0.238μg/ml,1.371±0.119μg/ml, 1.083±0.055μg/ml in a concentration dependent manner on A549/DDP cells. The difference compared with blank had statistical significance (P<0.05). The reversal fold was 5.066,7.254,9.178. The GST-πmRNA expression was down regulated after JS-K (0.1μM,0.3μM, 1μM) treated 48h. It induced concentration dependent GST-πmRNA expression changes on A549 and A549/DDP cells. The difference compared with blank had statistical significance (P< 0.05). Conclusion:JS-K can enhance the sensitivity of A549/DDP cells to cisplatin. The reversal effect may be relevant to the down regulation of GST-πmRNA expressions.更多还原