通过奥沙利铂抑制PI3K/Akt通路以增强TRAIL诱导结肠癌RKO细胞凋亡敏感性的实验研究  

Sensitivity Mechanism for Colon Cancer RKO Cell Apoptosis Induced by TRAIL Enhanced by the Inhibition of PI3K/Akt Pathway by Oxaliplatin

作　　者：徐明昊[1] 张悦[1] 孙明华[1] 李恩泽[2] 朱志图[2] 

机构地区：[1]辽宁医学院,辽宁省锦州市121000 [2]辽宁医学院附属第一医院

出　　处：《中国全科医学》2013年第18期2113-2116,共4页Chinese General Practice

摘　　要：目的通过亚毒性剂量奥沙利铂抑制PI3K/Akt通路以增强肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导结肠癌RKO细胞凋亡的敏感性,探讨其作用机制。方法将常规传代培养的人结肠癌RKO细胞分为空白对照组、对照组、TRAIL单药组(25、50、100、200、400 ng/ml)、奥沙利铂单药组(10、20、40、80μg/ml)及联合用药组(100 ng/ml TRAIL+40μg/ml奥沙利铂)。采用甲基偶氮唑蓝(MTT)检测对数生长期细胞,计算细胞增殖率和药物抑制细胞增殖50%的浓度(IC50);瑞氏-吉姆萨染色法观察细胞形态;流式细胞术检测细胞凋亡,计算细胞凋亡率;蛋白印迹法(Western blot)检测各组细胞蛋白变化。结果 TRAIL单药组24 h时细胞增殖率与对照组比较,差异均无统计学意义(P>0.05);奥沙利铂单药组24、48、72 h时IC50与对照组比较,差异均有统计学意义(P<0.05);联合用药组24 h时细胞增殖率与40μg/ml奥沙利铂单药组比较,差异有统计学意义(P<0.05)。100 ng/ml TRAIL组细胞凋亡率与对照组比较,差异无统计学意义(P>0.05);40μg/ml奥沙利铂组、联合用药组细胞凋亡率与对照组比较,差异均有统计学意义(P<0.05)。100 ng/ml TRAIL组p-Akt活化性增强,没有观察到活化的Caspase-3;40μg/ml奥沙利铂组和联合用药组p-Akt活化性被明显抑制,可以观察到活化的Caspase-3。对照组细胞形态正常,40μg/ml奥沙利铂组可见典型的凋亡小体。结论奥沙利铂可通过抑制PI3K/Akt通路活化而增强TRAIL诱导结肠癌RKO细胞凋亡的敏感性,为结肠癌的治疗提供了新思路。Objective To investigate the mechanism for colon cancer RKO cell apoptosis induced by TRAIL that is en-hanced by the inhibition of PDK/Akt pathway by oxaliplatin. Methods Sub - cultured RKO cells were divided into blank controlgroup, control group, single TRAIL group （25 , 50, 100, 200 and 400 ng/ml） , single oxaliplatin group （10, 20, 40 and80 jxg/ml） and drug combination group （ 100 ng/ml TRAIL ＋40 μg/ml oxaliplatin） . MTT assay was used to detect the log-arithm growth period and the cell proliferation rate and IC50 were calculated. Wright - Giemsa staining was used to observe morpho-logical changes. Flow cytometry was used to detect apoptosis and Western blot was used to detect protein expression. Results Thecell proliferation rate at 24 h in single TRAIL group and control group showed no statistically significant difference （ P 〉 0. 05 ）.The ICsq at 24 h, 48 h and 72 h in single oxaliplatin group and control group showed statistically significant differences （P 〈0. 05） . The cell proliferation rate at 24 h in drug combination group and 40 μg/ml single oxaliplatin group showed statisticallysignificant difference （ P 〈 0. 05 ） . The cell apoptosis rate in 100 ng/ml TRAIL group and control group showed no statisticallysignificant difference （ P 〉 0. 05 ） . Compared with control group, the cell apoptosis rate in 40 μg/ml oxaliplatin group, drugcombination group showed statistically significant differences （ P 〈 0. 05 ） . The activation of p - Akt was increased in 100 ng/mlTRAIL group and no activated Caspase - 3 was found. The activation of p - Akt was significantly inhibited in 40 jxg/ml oxaliplatingroup and drug combination group and activated Caspase - 3 was found. The cell morphology was normal in the control group andapoptosis body was found in 40 jxg/ml oxaliplatin group. Conclusion By inhibiting the activation of PI3K/Akt pathway, oxali-platin can enhance the sensitivity of TRAIL inducing the apoptosis of colon cancer RKO cells, which provides new ideas for th

关 键 词：TNF相关凋亡诱导配体 奥沙利铂 结肠肿瘤 PI3K Akt 细胞凋亡 

分 类 号：R735.35[医药卫生—肿瘤]