Rh-Apo2L联合长春瑞滨杀伤人肺腺癌A549细胞的作用研究  

The Killing Effect of Rh-Apo2L Combined with Vinorelbine on Human Lung Adenocarcinoma A549 Cells

作　　者：林宝钗[1] 陈金麟[2] 丁新华[2] 洪卫[1] 凌志强[1] 冯建国[1] 古翠萍[1] 洪丹[1] 胡林[2] 邵岚[2] 张沂平[1] 

机构地区：[1]浙江省肿瘤医院浙江省胸部肿瘤诊治技术研究重点实验室,浙江杭州310022 [2]浙江中医药大学,浙江杭州310053

出　　处：《肿瘤学杂志》2012年第3期161-165,共5页Journal of Chinese Oncology

基　　金：浙江省医药卫生科学研究基金资助项目(2007B025;2008A015;2010KYA032;2010KYA036);吴阶平医学基金(2011;320;6750.11059和2011;320;6750.11091)

摘　　要：[目的]探讨重组人凋亡素2配体(Rh-Apo2L)联合长春瑞滨(NVB)对人肺腺癌A549细胞的杀伤作用及机制。[方法]人肺腺癌A549细胞分成4组:对照组、Rh-Apo2L组、NVB组、联合组。用MTT法检测不同浓度Rh-Apo2L和NVB对A549细胞的抑制率,绘制细胞生长曲线,流式细胞仪测定细胞周期分布,RT-PCR方法检测死亡受体DR5基因mRNA的表达。[结果]MTT结果显示不同浓度Rh-Apo2L和NVB对人肺癌A549细胞的体外抑制作用均呈剂量效应关系,Rh-Apo2L处理48h后的IC20为3.48μg/ml、IC50为197.92μg/ml,NVB处理48h后的IC20为0.29μg/ml、IC50为3.44μg/ml。联合组对A549细胞的抑制作用高于单药组(P=0.001)。流式细胞分析显示含NVB药物两组在24hG2/M期比率高于对照组。3个实验组在24h、48h、72h后DR5基因mRNA相对表达量均较对照组多(P<0.05),但各实验组间均无统计学差异(P>0.05)。[结论]Rh-Apo2L与NVB联合应用能协同增强对人肺腺癌细胞株A549的体外抗肿瘤活性,其协同作用机制可能与细胞周期阻滞在G2/M期有关,但与DR5基因mRNA的表达无相关性。[Purpose] To investigate the killing effect of Rh-Apo2L in combination with vinorelbine（NVB） on human lung adenocarcinoma A549 cells and its mechanism.[Methods] Lung adenocarcinoma A549 cells were divided into four groups：the control group,Rh-Apo2L group,NVB group and the combination group.MTT assay was used to detect the inhibition of different concentrations of Rh-Apo2L and NVB to the A549 cells.Cell growth curves were drawn.Cell cycle distribution was determined by flow cytometry.The expression of death receptor DR5 mRNA was detected by RT-PCR.[Results] The MTT results showed that both Rh-Apo2L and NVB could inhibit the growth of A549 cells in a dose-dependent manner.After treatment with Rh-Apo2L for 48 h,the IC20 was 3.48 μg/ml and the IC50 was 197.92 μg /ml.After NVB treated for 48h,the IC20 was 0.29 μg/ml and the IC50 was 3.44 μg/ml.The inhibition rate of the combination group was higher than that in single drug group（P=0.001）.After treated for 24h,FCM showed that the ratio of cells in G2/M phase of the NVB-containing groups NVB group and the combination group were higher than that in the control group.Relative expression quantity of death receptor DR5 gene mRNA in three experimental groups was higher than that in control group（P0.05） in 24 h,48 h,72 h respectively,but no statistically significant differences were found between the experimental groups（P0.05）.[Conclusion] The combination of Rh-Apo2L and NVB could significantly enhance the antitumor activity to lung adenocarcinoma A549 cells in vitro.The combination mechanism may be related to the cell cycle arrest of G2/M phase,but not related to the expression of the DR5 gene mRNA.

关 键 词：Rh-Apo2L 细胞周期 肺肿瘤 长春瑞滨 

分 类 号：R734.2[医药卫生—肿瘤]