不同照射剂量联合吉非替尼对肺癌细胞系H358的影响

The effects of different dose combined gefitinib on lung cancer cell lines H358

目的探讨不同剂量单纯照射及照射联合吉非替尼对非小细胞肺癌细胞系H358存活率、增敏比及细胞凋亡、细胞周期的影响。方法体外培养肺腺癌细胞H358分为空白对照组、单纯照射组、单纯吉非替尼(Iressa)组、照射+Iressa组,照射剂量为0、2、4、6、8、12、16、20Gy,药物浓度为1μmol/L。通过细胞克隆形成实验,观察4组H358细胞存活情况,描绘细胞存活曲线;采用四噻唑比色试验(MTT)观察两组细胞生长受抑制情况,计算增敏比(SER);运用流式细胞仪检测细胞凋亡率和细胞周期。结果 (1)随着照射剂量增加细胞存活分数减小,单纯照射组单次剂量达到20Gy时,细胞克隆集落形成率为0,照射+Iressa组单次剂量达到16Gy时克隆集落形成率为0;(2)照射+Iressa组与单纯照射组相比,两组OD值差异有统计学意义(F剂量=62.644,P<0.001,F药物=233.572,P<0.001),两组OD值随着照射剂量的增高而减小,不同剂量之间OD值差异有统计学意义(F未加药=354.972,P<0.001;F加药=231.740,P<0.001),两组细胞放射增敏比(SER)与药物显著相关(P<0.001),照射+Iressa组SER较单纯照射组明显增高;(3)随着照射剂量的增加,凋亡率增高(P<0.001),H358细胞凋亡率与Iressa药物作用相关(P<0.001),单纯Iressa作用后细胞周期阻滞主要出现在G0/G1期,照射+Iressa组及单纯照射组主要出现G2/M期阻滞。结论增加单次照射剂量可降低H358细胞存活率,细胞凋亡与Iressa药物之间存在相关性,照射+Iressa能够增加细胞凋亡率,照射前使用Iressa能够增强照射对细胞的放射敏感性。

Objective This study aimed to explore the effects on the survival,sensitization enhancement ra-tio and cell apoptosis,cell cycle in non-small cell lung cancer cell lines H358 by a single different dose of irradiation and the irradiation combined with gefitinib.Methods Lung adenocarcinoma cells H358 in vitro were divided into four groups：control group,irradiation group,Iressa group,irradiation combined Iressa group,with dose 0-20 Gy and drug concentration 1μmol/L.Observe H358 cell survival by cell colony formation assay and depict cell survival curves;Observe cell growth inhibition by MTT and calculate SER;Detect cell apoptosis and cell cycle by flow cytometry;Process data by SPSS17.0.Results Cell colony for-mation assay showed that cell survival fraction decreased with the increase of irradiation dose,when a sin-gle dose irradiation reached 20Gy,cell clone formation rate was 0;in irradiation combined Iressa group, clonal colony formation rate was 0 when the dose reached 1 6 Gy.MTT results showed that by comparing Iressa combined irradiation group and simple irradiation group,there were significant differences between OD values in the two groups （Fdose=62.644,P〈0.001,Fdrug=233.572,P〈0.001）,two groups of OD val-ues decreased as the dose increased;OD value differences in different doses were statistically significant （Fnodrug=354.972,P〈0.001;Fdrug=231.740,P〈0.001）,two groups cell radiosensitization ratio （SER） were significantly associated with the drug （P 〈0.001）,SER of Iressa radiation group was significantly higher than that in the irradiation group;The result of flow cytometry showed that H358 cell apoptosis were associated with Iressa drug effects （P 〈0.001）,H358 cell apoptosis rate increased with the increase of single dose （P 〈0.001）.Simple Iressa group cell cycle arrest mainly appeared in G0/G1 phase,Iressa combined irradiation group and simple irradiation group mainly in G2/M phase.Conclusion Increased sin-gle dose decreased H358 cell survival;There was a correlation between cell apoptosis and the drug Iressa;Irradiation combined Iressa group increased the rate of cell apoptosis;Used Iressa before irradiation en-hanced the radiosensitivity of irradiation to cells.