CRABP I expression and the mediation of the sensitivity of human tumour cells to retinoic acid and irradiation
CRABP I expression and the mediation of the sensitivity of human tumour cells to retinoic acid and irradiation
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DOI：
10.1080/09553000310001632949
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发表时间：
2003-12-01
影响因子：
2.6
通讯作者：
Rodemann, HP
中科院分区：
文献类型：
--
作者：
Blaese, MA;Santo-Hoeltje, L;Rodemann, HP
目的：探讨细胞质维A酸结合蛋白1(CRABP I)和维甲酸受体β2(RAR-β2)在维甲酸对体外培养的人肿瘤细胞放射增敏中的作用。材料和方法：用维甲酸处理不同类型的人鳞癌细胞系，然后进行放射治疗。用集落形成实验检测药物治疗下的放射反应。用室温聚合酶链式反应和Western blotting检测不同处理条件下CRABP I、RAR-β和细胞周期蛋白D1mRNA和蛋白的表达水平。将CRABP I基因导入维甲酸敏感细胞株HTB35，观察CRABP I蛋白对维甲酸和辐射敏感性的调节作用，以及对RAR-b2和细胞周期蛋白D1表达的调控作用。结果：基础CRABP I水平与肿瘤细胞和正常成纤维细胞经维甲酸和电离辐射(IR)处理后的克隆存活率明显相关。与低基础表达的细胞相比，高基础表达CRABP I的细胞对联合维甲酸放射治疗的抵抗力更强。在维甲酸敏感的HTB35细胞中过表达CRABP I诱导了维甲酸不敏感的表型，抵抗维甲酸和辐射的联合处理。这种作用不依赖于RAR-b2的表达。CRABP I过表达可刺激细胞周期蛋白D1的表达，提示该细胞周期调控蛋白依赖于维甲酸代谢。结论：CRABP I不仅在维甲酸联合放射治疗后的维甲酸效应中起重要作用，而且在调节肿瘤细胞的放射敏感性中起重要作用。
Purpose: To examine the role cytoplasmic retinoic acid binding protein type 1 (CRABP I) and retinoic acid receptor beta2 (RAR-beta2) in mediating radiosensitization of human tumour cells in vitro by retinoic acid. Materials and methods: Human squamous cell carcinoma cell lines of different types were treated with retinoic acid followed by irradiation. Radiation response under drug treatment was detected by colony-formation assay. mRNA and protein expression levels of CRABP I, RAR-beta and cyclin D1 were investigated under different treatment conditions by room temperature polymerase chain reaction and Western blotting. The retinoic acid-sensitive cell line HTB35 was transfected for inducible CRABP I overexpression to test the role of this protein in modulating the sensitivity to retinoic acid and radiation as well as in regulating RAR-b2 and cyclin D1 expression. Results: The basal CRABP I level clearly correlated with the clonogenic survival of tumour cells and normal fibroblasts after treatment with retinoic acid and ionizing irradiation (IR). Cells expressing high basal CRABP I were more resistant to combined retinoic acid radiation treatment than cells with low basal expression. Overexpression of CRABP I in retinoic acid-sensitive HTB35 cells induced a retinoic acid-insensitive phenotype resistant to combined treatment with retinoic acid and radiation. This effect was independent of RAR-b2 expression. CRABP I overexpression resulted in stimulated cyclin D1 expression indicating the dependency of this cell cycle control protein on retinoic acid metabolism. Conclusion: CRABP I plays an important role not only in mediating the retinoid effects, but also in modulating the radiation sensitivity of tumour cells after combined retinoic acid radiation treatment.