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中国癌症防治杂志

中国癌症防治杂志 ›› 2023, Vol. 15 ›› Issue (2): 163-169.doi: 10.3969/j.issn.1674-5671.2023.02.08

• 基础研究 • 上一篇    下一篇

SLC1A5对三阴性乳腺癌MDA⁃MB⁃231细胞株放射敏感性的影响及作用机制

  

  1. 广西医科大学附属武鸣医院放疗科
  • 出版日期:2023-04-25 发布日期:2023-05-05
  • 通讯作者: 赵伟 E-mail：zwei20211@163.com
  • 基金资助:
    广西医疗卫生适宜技术开发与推广应用项目（S2018008）；广西中医药适宜技术开发与推广项目（GZSY22?70）；南宁市武鸣区科学研究与技术开发计划项目（20200213）

Effect of SLC1A5 on the radiosensitivity of triple negative breast cancer MDA-MB-231 cells and its mechanism

  • Online:2023-04-25 Published:2023-05-05

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摘要： 目的 探讨溶质载体家族1成员5（solute carrier family 1 member 5，SLC1A5）对三阴性乳腺癌MDA⁃MB⁃231细胞株放射敏感性的影响及其作用机制。方法 采用直线加速器X射线和SLC1A5抑制剂L⁃γ⁃谷氨酰⁃对⁃硝基苯胺（L⁃γ⁃glutamyl⁃p⁃nitroanilide，GPNA）处理三阴性乳腺癌MDA⁃MB⁃231细胞株，选取4 Gy、8 Gy剂量单独照射或分别联合3 mmol/L GPNA处理，采用CCK⁃8法检测细胞增殖能力；然后选取8 Gy剂量单独照射或联合3 mmol/L GPNA进行处理，分为对照组（DMSO组）、SLC1A5抑制剂组（GPNA组）、照射组（IR组）、联合组（IR+GPNA组）。采用细胞克隆形成实验、划痕实验、Transwell小室实验分别检测细胞克隆形成能力、迁移能力和侵袭能力。采用活性氧（reactive oxygen species，ROS）荧光探针（DCFH⁃DA）检测细胞内的ROS水平，丙二醛（malondialdehyde，MDA）试剂盒检测细胞内的MDA含量，Mito⁃Tracker Red CMXRos染色实验观察细胞线粒体形态，RT⁃qPCR法检测前列素内环氧化物合成酶2（prostaglandin synthase 2，PTGS2）基因的表达水平。结果 与DMSO组相比，4 Gy组、8 Gy组、4 Gy+GPNA组和8 Gy+GPNA组细胞增殖能力均下降（均P<0.01），且4 Gy+GPNA组、8 Gy+GPNA组细胞增殖能力均低于对应的单纯照射组（均P<0.01），其中高辐射剂量组下降更明显；与DMSO组相比，IR组、GPNA组和IR+GPNA组的细胞克隆形成能力、迁移能力、侵袭能力均降低（均P<0.01），且IR+GPNA组降低更明显。与DMSO组相比，GPNA组、IR组和IR+GPNA组都存在不同程度的细胞内ROS水平增加，线粒体形态缩小，MDA含量升高以及PTGS2基因表达升高（均P<0.05），其中IR+GPNA组变化更为明显。结论 SLC1A5抑制剂在照射的基础上进一步抑制三阴性乳腺癌MDA⁃MB⁃231细胞的增殖、克隆形成、迁移及侵袭能力，提高细胞放射敏感性，其机制可能与铁死亡激活有关。

关键词: 乳腺癌, MDA-MB-231细胞, 放射敏感性, SLC1A5, 铁死亡

Abstract: Objective To investigate the effect of solute carrier family 1 member 5 (SLC1A5) on the radiosensitivity of triple⁃negative breast cancer MDA⁃MB⁃231 cell lines and its mechanism. Methods The triple⁃negative breast cancer MDA⁃MB⁃231 cell lines  were treated with X⁃ray and SLC1A5 inhibitor L⁃γ⁃glutamyl⁃p⁃nitroanilide (GPNA). The cells were irradiated with 4 Gy or 8 Gy alone or combined with 3 mmol/L GPNA, and the proliferation ability of cells was detected by CCK⁃8 method. Then the cells treated with 8 Gy irradiation alone or combined with 3 mmol/L GPNA were divided into control (DMSO) group, SLC1A5 inhibitor (GPNA) group, irradiation (IR) group, and combination (IR+GPNA) group. Clone formation test, Scratch assay and Transwell assay were used to detect the clonogenesis ability, migration ability and the invasion ability of cells, respectively. Reactive oxygen species (ROS) fluorescent probe (DCFH⁃DA) was used to detect the contents of ROS in cells. Malondialdehyde (MDA) kits were used to detect the contents of MDA in cells. Mito⁃tracker Red CMXRos staining assay was used to observe mitochondrial morphology. The expression level of PTGS2 gene was detected by RT⁃qPCR. Results Compared with the DMSO group, the cell proliferation ability decreased in the 4 Gy, 8 Gy, 4 Gy+GPNA and 8 Gy+GPNA groups(all P<0.01), the proliferation ability of cells in the 4 Gy+GPNA and 8 Gy+GPNA groups was lower than that of the corresponding simple irradiation groups(all P<0.01), and the decrease was more obvious in the high radiation dose group. Compared with the DMSO group, the cell clone formation ability, migration ability and invasion ability were reduced in the IR, GPNA and IR+GPNA groups, and the reduction was more significant in the IR+GPNA group. Compared with the DMSO group, the GPNA, IR and IR+GPNA groups had increased ROS levels, MDA contents and PTGS2 gene expression (all P<0.05) and reduced mitochondrial morphology (P<0.01), and such changes were most pronounced in the IR+GPNA group. Conclusions SLC1A5 inhibitor can further inhibit the proliferation. clonogenesis migration and invasion, of triple⁃negative breast cancer MDA⁃MB⁃231 cells on the basis of irradiation, and improve the radiosensitivity of cells. The underlying mechanism may be related to the activation of ferroptosis.

Key words: Breast cancer, MDA-MB-231 cells, Radiosensitivity, SLC1A5, Ferroptosis 

中图分类号: 

  • R737.9