谷氨酰胺对结直肠癌HT-29细胞放射敏感性的影响及机制探讨  

Effect and underlying mechanism of glutamine on radiosensitivity of colon cancer cells

作　　者：卢亨 倪向敏 于生财 梁馨予 朱文艺 李忠俊 王建 LU Heng;NI Xiangmin;YU Shengcai;LIANG Xinyu;ZHU Wenyi;LI Zhongjun;WANG Jian(Department of Nutrition,Second Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400037,China;Department of Blood Transfusion,Second Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,400037,China)

机构地区：[1]陆军军医大学(第三军医大学)第二附属医院营养科,重庆1400037 [2]陆军军医大学(第三军医大学)第二附属医院输血科,重庆1400037

出　　处：《陆军军医大学学报》2024年第9期1007-1014,共8页Journal of Army Medical University

基　　金：重庆英才·创新领军人才(医学领域)计划项目(CQYC20220303514)。

摘　　要：目的观察不同浓度谷氨酰胺(glutamine,Gln)对结直肠癌HT-29细胞放射敏感性的影响,并探讨其可能机制。方法体外培养HT-29细胞,将对数生长期的HT-29细胞按照Gln不同浓度设置对照组(2 mmol/L,培养基基础浓度)、实验组Ⅰ(4 mmol/L)、实验组Ⅱ(6 mmol/L)、实验组Ⅲ(8 mmol/L),预处理2 h后给予Co 60源放射。通过CCK-8法检测未放射与放射后不同浓度Gln对细胞活力的影响;克隆形成实验检测放射后不同浓度Gln对细胞放射敏感性的影响;活性氧(ROS)试剂盒检测放射后24 h细胞ROS水平;流式细胞仪检测放射后48 h细胞凋亡率;蛋白印迹法测定放射后各组细胞Nrf2、HO-1、cleaved-Caspase3蛋白的表达水平。结果未放射的HT-29细胞在Gln干预24 h后实验组Ⅱ、实验组Ⅲ细胞活力明显高于对照组、实验组Ⅰ(P<0.05)。HT-29细胞放射24 h后,各实验组细胞活力明显高于对照组(P<0.05);放射后14 d,各实验组细胞克隆形成数均高于对照组(P<0.05);放射后24 h,各实验组ROS水平均低于对照组(P<0.05);放射后48 h,各实验组细胞凋亡率均低于对照组(P<0.05)。实验组ⅠNrf2表达高于对照组(P<0.05);实验组Ⅱ、ⅢNrf2、HO-1表达均高于对照组和实验组Ⅰ(P<0.05),cleaved-Caspase3表达均低于对照组和实验组Ⅰ(P<0.05);实验组Ⅲcleaved-Caspase3表达低于实验组Ⅱ(P<0.05)。结论谷氨酰胺能显著降低HT-29细胞的放射敏感性,其机制与减轻氧化应激损伤和减少细胞凋亡有关,这可能不利于结直肠癌患者放疗。Objective To observe the effect of different concentrations of glutamine(Gln)on the radiosensitivity of colorectal cancer HT-29 cells and explore the possible mechanism.Methods According to different Gln concentrations,HT-29 cells at logarithmical growth were divided into control group(2 mmol/L,as the basal medium concentration group)and experimental groupsⅠ,ⅡandⅢ(4,6 and 8 mmol/L).After a 2-hour pre-treatment,all groups were exposed to 8 Gy irradiation of a Co-60 radiation source.CCK-8 assay and clonal formation assay were used respectively to explore the effects of different Gln concentrations on cell viability and cell radiosensitivity after irradiation.The level of reactive oxygen species(ROS)in each group was measured in 24 h after irradiation,and the apoptotic rate was detected with flow cytometry in 48 h after irradiation.The protein expression levels of Nrf2,HO-1,and cleaved-Caspase3 were determined by Western blotting.Results In 24 h after Gln intervention,the cell viability of experimental groupsⅡandⅢof non-irradiated HT-29 cells was significantly higher than that of the control group and of experimental groupⅠ(P<0.05).In 24 h after radiation,the cell viability of each experimental group was significantly higher than that of the control group(P<0.05).In 14 d after radiation,there were more clone formation in each experimental group than the control group(P<0.05).The ROS level was significantly lower in each experimental group than the control group in 24 h after radiation(P<0.05).After 48 h of radiation,the apoptotic rate was notably lower in each experimental group than the control group(P<0.05).The expression level of Nrf2 in the experimental groupⅠwas higher than that of the control group(P<0.05),those of Nrf2 and HO-1 in the experimental groupsⅡandⅢwere higher than those of the control group and experimental groupⅠ(P<0.05).While the expression of cleaved-Caspase3 in the experimental groupsⅡandⅢwas lower than the control group and experimental groupⅠ(P<0.05),and it in the ex

关 键 词：谷氨酰胺 HT-29细胞 氧化应激损伤 细胞凋亡 放射敏感性 

分 类 号：R730.55[医药卫生—肿瘤] R735.35[医药卫生—临床医学] R977.4