[1]贺戈,汪国建,赵娜,等.柳氮磺吡啶拮抗电离辐射对乳腺癌转移促进作用的实验研究 [J].陆军军医大学学报(原第三军医大学学报),2022,44(18):1797-1808.
　HE Ge,WANG Guojian,ZHAO Na,et al.Sulfasalazine antagonizes boosted metastasis of breast cancer induced by ionizing radiation[J].J Amry Med Univ (J Third Mil Med Univ),2022,44(18):1797-1808.
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柳氮磺吡啶拮抗电离辐射对乳腺癌转移促进作用的实验研究
 
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陆军军医大学学报（原第三军医大学学报）[ISSN:1000-5404/CN:51-1095/R]

卷:
44卷
期数:
2022年第18期
页码:
1797-1808
栏目:
基础医学
出版日期:
2022-09-30

文章信息/Info

Title:
Sulfasalazine antagonizes boosted metastasis of breast cancer induced by ionizing radiation
作者:
贺戈; 汪国建; 赵娜; 龙爽; 王涛; 明佳
重庆医科大学附属第二医院乳甲外科；陆军军医大学（第三军医大学）军事预防医学系防原医学教研室，全军复合伤研究所，创伤、烧伤与复合伤国家重点实验室，重庆市纳米医学工程研究中心
Author(s):
HE Ge1;  WANG Guojian2;  ZHAO Na2;  LONG Shuang2;  WANG Tao2;  MING Jia1

1Department of Breast and Thyroid Surgery, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010; 2State Key Laboratory of Trauma, Burns and Combined Injury, Department of Anti-Radiation Medicine, Chongqing Engineering Research Center for Nanomedicine, Institute of Combined Injury, Faculty of Military Preventive Medicine, Army Medical University (Third Military Medical University), Chongqing, 400038, China

关键词:
乳腺癌; 转移; 电离辐射; 上皮-间质转化; 柳氮磺吡啶
Keywords:
breast cancer;  metastasis;  ionizing radiation;  epithelial-mesenchymal transition;  sulfasalazine 
分类号:
R737.9; R966; R979.1
文献标志码:
A
摘要:
目的评价柳氮磺吡啶（sulfasalazine，SSZ）对电离辐射（ionizing radiation，IR）诱导离体乳腺癌细胞上皮-间质转化（epithelial-to-mesenchymal transition, EMT）、迁移、侵袭等转移相关表型的抑制作用，探讨其作为治疗措施对在体局部IR促进乳腺癌肺转移的拮抗作用。方法以体外培养两种乳腺癌细胞株（MCF-7、4T1）为研究对象，采用不同剂量（2、4、6、8、10 Gy）X射线照射后观察细胞形态改变，Western blot检测Vimentin、Snail等EMT相关标志物的表达变化。选取
4 Gy X射线照射细胞，评价SSZ（0.5 mmol/L）对IR后细胞EMT、迁移、侵袭等转移相关表型的影响，并检测MMP2/9和xCT等蛋白的表达。在BALB/c雌鼠建立4T1细胞乳腺癌肺转移模型，实验分4组：Con组、SSZ组、IR组和IR+SSZ组。IR采用局部X射线（4 Gy）照射；SSZ采用腹腔给药8 mg/只，2次/d；IR+SSZ组采用局部X射线（4 Gy）照射后，以腹腔给药SSZ 8 mg/只，2 次 /d。荷瘤30 d后取材，计算原位瘤大小和肺转移结节数目，HE染色观察原位瘤和肺转移灶情况。结果体外培养乳腺癌细胞在IR后出现明显的EMT形态改变，Vimentin、Snail等EMT相关蛋白表达上调，呈一定剂量相关性。SSZ能够抑制IR诱导的乳腺癌细胞EMT形态和标志蛋白改变，抑制IR诱导的乳腺癌细胞迁移和侵袭能力增强的表型（P<0.05），下调IR诱导的MMP2/9、xCT等蛋白增强表达。在体动物模型与离体细胞结果相对应，IR可以显著抑制原位瘤生长，与Con组比较，IR组[（1.24±0.32）cm3]和IR+SSZ组[（0.83±0.54）cm3]的小鼠原位瘤平均体积显著降低（P<0.05），且IR+SSZ组较IR组降低更显著（P<0.05）；IR组小鼠肺转移结节数为[（19.00±8.31）个]，较Con组[（8.67±7.88）个]显著增多（P<0.05），IR+SSZ组小鼠肺转移结节数[（6.10±7.78）个]较IR组显著降低（P<0.05），而单纯SSZ组小鼠肺转移结节数[（4.30±5.39）个]与Con组相比差异无统计学意义。HE染色结果显示，IR组肺转移灶数量增多，面积增加，边界欠清，有明显弥散的趋势；而IR+SSZ组肺转移灶面积较小，边界较清晰。结论SSZ能有效抑制IR诱导乳腺癌细胞EMT、迁移、侵袭，并显著拮抗在体局部照射对乳腺癌肺转移的促进作用。
Abstract:

ObjectiveTo evaluate the inhibitory effects of sulfasalazine (SSZ) on ionizing radiation (IR) induced epithelial-to-mesenchymal transition (EMT), migration, invasion, and other metastasis-related phenotypes in breast cancer cells in vitro, and to investigate its antagonistic effects as a therapeutic measure against breast cancer lung metastasis promoted by in vivo local IR. MethodsCultured breast cancer cell lines MCF-7 and 4T1 were used to study the morphological changes of cells after different doses of X-ray irradiation (2, 4, 6, 8 and 10 Gy) and the expression changes of EMT-related markers such as Vimentin and Snail were detected by Western blotting. A cell model with 4 Gy of X-ray irradiation was selected to evaluate the effect of SSZ (0.5 mmol/L) on metastasis-related phenotypes such as EMT, migration, and invasion of cells after IR, and on the protein expression of MMP2/9 and xCT after IR. After xenograft model of 4T1 cell breast cancer lung metastasis was established in BALB/c female mice, the mice were divided into control group, SSZ group (intraperitoneal injection of 8 mg SSZ, 2 times per day), IR group (X-ray 4 Gy irradiation) and IR+SSZ group （X-ray 4 Gy irradiation, intraperitoneal injection of 8 mg SSZ, 2 times per day）. The tumor masses were harvested in 30 d after tumor loading, and the size of in situ tumor and the number of lung metastatic nodes were measured and calculated, and HE staining was used to observed in situ tumor and lung metastases. ResultsCultured breast cancer cells showed significant EMT morphological alterations after IR, and the expression of Vimentin and Snail was upregulated in a dose-related manner. SSZ inhibited IR-induced changes in EMT morphology and marker proteins, suppressed IR-induced phenotypes of enhanced migration and invasion of breast cancer cells (P<0.05), and down-regulated IR-induced enhanced expression of MMP2/9, xCT, and other proteins. In vivo animal model corresponded to the ex vivo cellular results that IR significantly inhibited in situ tumor growth. Compared with the control mice (1.70±0.54 cm3), the mean tumor volume was significantly smaller in the IR group (1.24±0.32 cm3) and IR+SSZ group (0.83±0.54 cm3) (P<0.05), and the reduction was more significant in the latter than in the former group (P<0.05). The number of lung metastases in the IR group (19.00±8.31) was significantly higher than that in the Con group (8.67±7.88) (P<0.05), and the number in the IR+SSZ group (6.10±7.78) was significantly lower than the IR group (P<0.05), while the number in the SSZ group (4.30±5.39) was not statistically different from the Con group. HE pathology showed that the number of metastases in the IR group was increased, the area was enlarged, the border was not clear, and the metastases had a tendency to diffuse. In contrast, the lung metastases in the IR+SSZ group had clearer borders and a weaker tendency to invade outward. ConclusionSSZ effectively inhibits IR-induced EMT, migration, and invasion of breast cancer cells and significantly antagonizes the lung metastasis-promoting effects of in vivo local irradiation of breast cancer. 

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更新日期/Last Update: 2022-09-23