全反式维甲酸对食管癌EC9706细胞系化疗敏感性的影响

Promoting effect of all-trans retinoic acid on the chemosensitivity of esophageal cancer EC9706 cells

目的 探讨全反式维甲酸（ATRA）对食管癌EC9706细胞系化疗敏感性的影响及其作用机制.方法 将常规传代培养的食管癌EC9706细胞分为4组,即ATRA作用组（加入终浓度为1μmol/L的ATRA）、5-氟尿嘧啶（5＋Fu）作用组（加入终浓度为50mg/L的5-Fu）、联合用药组（加入终浓度为1μmol/L的ATRA和终浓度为5omg/L的5-Fu）和空白对照组（仅含EC9706细胞）.采用原位酶末端标记（TUNEL）法检测各组细胞的凋亡情况,采用流式细胞仪检测各组细胞的凋亡率和细胞周期的变化.结果 TUNEL法检测结果显示,联合用药组的凋亡率达89.7%,明显高于ATRA作用组（38.3%）和5-Fu作用组（40.3%,均P＜0.05）.流式细胞仪检测结果显示,联合用药组的凋亡率达76.9%±2.7%,明显高于ATRA作用组（38.2%±2. 6%）和5-Fu作用组（45.2%±2.3%,均p＜0.05）.联合用药组G1期细胞比例明显增加,达83.4%±3.0%,明显高于ATRA作用组（48.2%±2.5%）和5-Fu作用组（53.2%±2.6%,均P＜0.05）;联合用药组S和G2/M期细胞比例明显下降,与其他各组比较,差异均有统计学意义（均P＜0.05）.ATRA作用组与5-Fu作用组的凋亡率和各期细胞比例的差异均无统计学意义（均P＞0.05）.结论 ATRA有增加食管癌EC97O6细胞对5-Fu敏感性的作用,其作用机制与其调节细胞周期的变化和增加细胞的凋亡有关.

Objective To investigate the impact of all-trans retinoic acid （ATRA） on chcmosensitivity to esophageal squamous cell carcinoma EC9706 cells in vitro and its mechenism. Methods EC9706 cells were routinely cultured as the control group. The experimental group was divided into three groups.The ATRA group with ATRA in final concentration of 1 μmol/L; the 5-Fu group with 5 -Fu in final concentrationof 50 mg/L; the combined treatment group with ATRA in final concentration of 1 μmol/L and 5-Fu 50 mg/L.The cell apoptosis was detected by terminal deexynucleotidy transferase mediated dUTP nick end labelling （TUNEL）The cell cyele and apoptosis were detected by flow cytometry Results The results of TUNFL showed that m the combined treatment group appeared a large number of apoptotic cells, and their nuclei were stained brown, with a positive rate of 89.7%. There was a significant difference in the comparison with the ATRA group （38. 3%） and 5-Fu group （40. 3%o） （P 〈 0. 05） The flow cytometry showed that the ATRA ＋ 5-Fu group had a significantly higher apoptosis rate （76.9% ±2. 7%） than that in the ATRA group （38.2% ±2.6%） and 5-Fu group （45.2% ±2. 3%） （P 〈0.05） The ratio of cells in G1 phase increased in the ATRA＋ 5-Fu group （83.4%±3.0%）, significantly higher than（48.2% ±2.5%）in the ATRA group and （53.2% ± 2. 6%） in the 5-Fu group （P 〈 0. 05） The ratio of cells in S＋G2/M phase was decreaced in the ATRA ＋ 5-Fu group, with a significant difference （P 〈 0.05） when compared with other groups. There was no significant difference between the ATRA group and 5-Fu group （P 〉 0.05）in the apoptosis rate and the proportion of cells at different phases. Conclusion ATRA can induce apoptosis of esophageal carcinoma EC9706 cells in vitro. The combination of ATRA and 5-Fu may enhancethe chemotherapeutic efficacy.