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

卢亨; 倪向敏; 于生财; 梁馨予; 朱文艺; 李忠俊; 王建

doi:10.16016/j.2097-0927.202308077

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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

基础医学 | 更新时间：2026-01-09

- 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
- 陆军军医大学学报 2024年46卷第9期页码：1007-1014
- 作者机构：
  
  1.400037 重庆，陆军军医大学(第三军医大学)第二附属医院：营养科
  2.400037 重庆，陆军军医大学(第三军医大学)第二附属医院：输血科
- 作者简介：
  
  [ 王建，E-mail：wangjian_1996@aliyun.com WANG Jian, E-mail: wangjian_1996@aliyun.com
- 基金信息：
  
  重庆英才·创新领军人才(医学领域)计划项目(CQYC20220303514)
- DOI：10.16016/j.2097-0927.202308077
  中图分类号： R730.55;R735.35;R977.4
- 收稿：2023-08-19，
  
  修回：2024-01-08，
  
  纸质出版：2024-05-15
- 稿件说明：
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卢亨, 倪向敏, 于生财, 等. 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[J]. 陆军军医大学学报, 2024,46(9):1007-1014.

Heng LU, Xiangmin NI, Shengcai YU, et al. Effect and underlying mechanism of glutamine on radiosensitivity of colon cancer cells[J]. Journal of Army Medical University, 2024, 46(9): 1007-1014.
卢亨, 倪向敏, 于生财, 等. 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[J]. 陆军军医大学学报, 2024,46(9):1007-1014. DOI： 10.16016/j.2097-0927.202308077.

Heng LU, Xiangmin NI, Shengcai YU, et al. Effect and underlying mechanism of glutamine on radiosensitivity of colon cancer cells[J]. Journal of Army Medical University, 2024, 46(9): 1007-1014. DOI： 10.16016/j.2097-0927.202308077.

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(

＜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(

＜0.05)；放射后14 d，各实验组细胞克隆形成数均高于对照组(

＜0.05)；放射后24 h，各实验组ROS水平均低于对照组(

＜0.05)；放射后48 h，各实验组细胞凋亡率均低于对照组(

＜0.05)。实验组ⅠNrf2表达高于对照组(

＜0.05)；实验组Ⅱ、Ⅲ Nrf2、HO-1表达均高于对照组和实验组Ⅰ(

＜0.05)，cleaved-Caspase3表达均低于对照组和实验组Ⅰ(

＜0.05)；实验组Ⅲ cleaved-Caspase3表达低于实验组Ⅱ(

＜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。

Abstract

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 groups Ⅰ

Ⅱ and Ⅲ (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 groups Ⅱ and Ⅲ of non-irradiated HT-29 cells was significantly higher than that of the control group and of experimental group Ⅰ (

0.05). In 24 h after radiation

the cell viability of each experimental group was significantly higher than that of the control group (

0.05). In 14 d after radiation

there were more clone formation in each experimental group than the control group (

0.05). The ROS level was significantly lower in each experimental group than the control group in 24 h after radiation (

0.05). After 48 h of radiation

the apoptotic rate was notably lower in each experimental group than the control group (

0.05). The expression level of Nrf2 in the experimental group Ⅰ was higher than that of the control group (

0.05)

those of Nrf2 and HO-1 in the experimental groups Ⅱ and Ⅲ were higher than those of the control group and experimental group Ⅰ (

0.05). While the expression of cleaved-Caspase3 in the experimental groups Ⅱ and Ⅲ was lower than the control group and experimental group Ⅰ (

0.05)

and it in the experimental group Ⅲ was lower than that of experimental group Ⅱ (

0.05).

Conclusion

Gln can significantly reduce the radiosensitivity of HT-29 cells

which is associated with its reducing oxidative stress damage and reducing cell apoptosis. Our results suggest that Gln might be detrimental to radiation therapy in patients with colorectal cancer.

关键词

Keywords

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