国际肿瘤学杂志 ›› 2023, Vol. 50 ›› Issue (8): 457-462.doi: 10.3760/cma.j.cn371439-20230420-00088

• 论著 • 上一篇    下一篇

福瑞替尼对三阴性乳腺癌血管生成、肿瘤生长及IRE1-ASK1-JNK通路的影响

潘书兰, 刘畅(), 贺平   

  1. 哈励逊国际和平医院肿瘤科,衡水 053000
  • 收稿日期:2023-04-20 修回日期:2023-06-13 出版日期:2023-08-08 发布日期:2023-10-24
  • 通讯作者: 刘畅 E-mail:952839102@qq.com

Effect of fritinib on angiogenesis, tumor growth and IRE1-ASK1-JNK pathway in triple negative breast cancer

Pan Shulan, Liu Chang(), He Ping   

  1. Department of Oncology, Harrison International Peace Hospital, Hengshui 053000, China
  • Received:2023-04-20 Revised:2023-06-13 Online:2023-08-08 Published:2023-10-24
  • Contact: Liu Chang E-mail:952839102@qq.com

摘要: 目的 探讨福瑞替尼对三阴性乳腺癌血管生成、肿瘤生长及跨膜蛋白肌醇需求酶1(IRE1)-细胞凋亡信号调节激酶1(ASK1)-c-Jun氨基端激酶(JNK)通路的影响。方法 取三阴性乳腺癌细胞株MDA-MB-231,将其分为生理盐水(NS)组、低剂量福瑞替尼(LD)组、中剂量福瑞替尼(MD)组、高剂量福瑞替尼(HD)组,NS组加入100 μmol/L的生理盐水,LD组、MD组、HD组分别加入25、50、100 μmol/L的福瑞替尼。MTT法检测细胞增殖,流式细胞仪检测细胞凋亡,细胞三维培养观察细胞形成拟态血管能力,蛋白质印迹法检测血管生成及IRE1-ASK1-JNK通路相关指标。将20只三阴性乳腺癌大鼠模型采用随机数字表法分为对照组、实验组,每组10只,对照组给予生理盐水灌胃,实验组给予100 μmol/L福瑞替尼灌胃,观察并记录两组大鼠肿瘤瘤体生长情况。 结果 NS组、LD组、MD组、HD组的48 h细胞增殖率分别为(85.44±5.58)%、(73.24±4.95)%、(61.53±4.07)%、(50.23±2.97)%(F=4.01,P=0.002),与NS组相比,LD组、MD组、HD组细胞增殖率显著降低,且呈剂量依赖性(均P<0.05);NS组、LD组、MD组、HD组的细胞凋亡率分别为(3.41±0.39)%、(18.75±1.94)%、(24.97±2.58)%、(38.62±3.27)%(F=18.99,P<0.001),与NS组相比,LD组、MD组、HD组细胞凋亡率显著升高,且呈剂量依赖性(均P<0.05);NS组、LD组、MD组、HD组的拟态血管数目分别为19.58±2.11、15.67±2.02、11.57±1.73、5.20±1.23(F=3.28,P=0.008),与NS组相比,LD组、MD组、HD组拟态血管数目显著降低,且呈剂量依赖性(均P<0.05);NS组、LD组、MD组、HD组的血管内皮生长因子(VEGF)蛋白相对表达量分别为2.36±0.21、1.79±0.17、1.48±0.14、0.94±0.10(F=5.17,P<0.001),IRE1蛋白相对表达量分别为1.18±0.12、1.67±0.18、2.03±0.24、2.39±0.28(F=5.55,P<0.001),ASK1蛋白相对表达量分别为1.09±0.11、1.46±0.13、1.81±0.18、2.33±0.21(F=5.32,P<0.001),JNK蛋白表达分别为1.01±0.09、1.48±0.14、1.86±0.21、2.28±0.24(F=6.92,P<0.001),与NS组相比,LD组、MD组、HD组细胞VEGF蛋白表达显著降低,IRE1、ASK1、JNK蛋白表达显著升高,且呈剂量依赖性(均P<0.05);与对照组相比,实验组瘤体质量、体积明显降低[(0.55±0.10)g比(1.37±0.15)g,t=14.38,P<0.001;(77.39±3.21)mm3比(118.26±5.34)mm3t=20.74,P<0.001],抑瘤率显著升高[(71.23±3.85)%比(32.56±3.08)%,t=24.80,P<0.001]。结论 福瑞替尼对三阴性乳腺癌细胞活性具有抑制作用,可显著减少其血管生成,抑制肿瘤生长,可能与激活IRE1-ASK1-JNK通路相关。

关键词: 三阴性乳腺癌, 福瑞替尼, 血管生成, 肿瘤生长, IRE1-ASK1-JNK通路

Abstract: Objective To investigate the effects of fritinib on angiogenesis, tumor growth and inositol requiring enzyme 1 (IRE1)-apoptosis signal regulating kinase 1 (ASK1)-c-Jun N-terminal kinase (JNK) pathway in triple negative breast cancer. Methods Triple negative breast cancer cells MDA-MB-231 were taken and divided into normal saline (NS) group, low-dose fritinib (LD) group, medium-dose fritinib (MD) group and high-dose fritinib (HD) group. NS group was added with 100 μmol/L normal saline. LD group, MD group and HD group were added with 25, 50 and 100 μmol/L fritinib, respectively. Cell proliferation was detected by MTT assay, cell apoptosis was detected by flow cytometry, the ability of cells to form mimicry vessels was observed by three-dimensional cell culture, and the related indexes of angiogenesis and IRE1-ASK1-JNK pathway were detected by Western blotting. Twenty triple-negative breast cancer rat models were divided into control group and experimental group by random number table method, with 10 rats in each group. The control group was given normal saline gavage and the experimental group was given 100 μmol/L fritinib gavage. The tumor growth of rats in the two groups was observed and recorded. Results The 48 h cell proliferation rates of NS group, LD group, MD group and HD group were (85.44±5.58)%, (73.24±4.95)%, (61.53±4.07)% and (50.23±2.97)%, respectively (F=4.01, P=0.002). Compared with the NS group, the cell proliferation rate in LD, MD and HD groups was significantly decreased in a dose-dependent manner (all P<0.05). The apoptosis rates of NS group, LD group, MD group and HD group were (3.41±0.39)%, (18.75±1.94)%, (24.97±2.58)% and (38.62±3.27)%, respectively (F=18.99, P<0.001). Compared with the NS group, the apoptosis rate of LD, MD and HD groups was significantly increased in a dose-dependent manner (all P<0.05). The number of mimicry vessels in NS group, LD group, MD group and HD group was 19.58±2.11, 15.67±2.02, 11.57±1.73 and 5.20±1.23, respectively (F=3.28, P=0.008). Compared with the NS group, the number of mimicry vessels in LD group, MD group and HD group was significantly reduced. The results were dose-dependent (all P<0.05). vascular endothelial growth factor (VEGF) protein expression in NS group, LD group, MD group and HD group was 2.36±0.21, 1.79±0.17, 1.48±0.14 and 0.94±0.10, respectively (F=5.17, P<0.001). The expression of IRE1 protein was 1.18±0.12, 1.67±0.18, 2.03±0.24 and 2.39±0.28, respectively (F=5.55, P<0.001). The expression of ASK1 protein was 1.09±0.11, 1.46±0.13, 1.81±0.18, 2.33±0.21 (F=5.32, P<0.001), respectively. JNK protein expression was 1.01±0.09, 1.48±0.14, 1.86±0.21 and 2.28±0.24, respectively (F=6.92, P<0.001). Compared with the NS group, VEGF protein expression in LD, MD and HD groups was significantly decreased, and the expressions of IRE1, ASK1 and JNK were significantly increased in a dose-dependent manner (all P<0.05). Compared with the control group, the tumor weight and volume of the experimental group were significantly decreased [(0.55±0.10)g vs. (1.37±0.15)g, t=14.38, P<0.001; (77.39±3.21)mm3 vs. (118.26±5.34)mm3, t=20.74, P<0.001], tumor inhibition rate was significantly increased [(71.23±3.85)% vs. (32.56±3.08)%, t=24.80, P<0.001]. Conclusion Fritinib has an inhibitory effect on the activity of triple negative breast cancer cells, which can significantly reduce their angiogenesis and inhibit tumor growth. Moreover, it is related to the activation of IRE1-ASK1-JNK pathway.

Key words: Triple negative breast neoplasms, Fritinib, Angiogenesis, Tumor growth, IRE1-ASK1-JNK path