van der Leede B M, van den Brink C E, van der Saag P T
Hubrecht Laboratory, Netherlands Institute for Developmental Biology, Utrecht, The Netherlands.
Mol Carcinog. 1993;8(2):112-22. doi: 10.1002/mc.2940080208.
Retinoic acid (RA) has profound effects on cell proliferation and differentiation both in vitro and in vivo. Many human cell lines are known to be sensitive to the growth-inhibitory action of RA. We analyzed established human solid tumor-derived cell lines for their RA sensitivity. Growth inhibition by RA in monolayer was examined by [3H]thymidine incorporation and cell proliferation. Here we report that 11 widely used human cell lines were RA resistant. The majority are carcinoma derived (A-431, BT-20, C-41, ACHN, HCT116, 293, A549, and PA-1); two are sarcoma derived (Saos-2 and A673); and one is a melanoma cell line (A-375). Since nuclear retinoid receptors are implicated in the biological effects of RA, we examined the expression of retinoic acid receptors (RARs) RAR alpha, RAR beta, RAR gamma, and the retinoid X receptors (RXRs) RXR alpha, RXR beta, and RXR gamma in the RA-resistant cell lines by northern blotting and by RNase protection analysis for RAR beta. RAR alpha transcripts were constitutively expressed in all cell lines. By contrast, RAR beta was expressed in only seven RA-resistant cell lines (Saos-2, ACHN, 293, A549, A-375, A673, and PA-1), and its level was enhanced by RA in some cases. In most cell lines, RAR gamma expression was low and was not affected by RA. The RXR genes showed a very distinct expression pattern in the group of selected cell lines. In general, RXR alpha was the most abundantly expressed subtype, RXR beta was expressed at low levels, and RXR gamma could not be detected. In none of the RA-resistant cell lines was RXR expression modulated by RA. The results presented here indicate that the resistance of these human tumor cell lines to RA cannot be simply correlated with expression of RAR or RXR or both.
维甲酸(RA)在体外和体内对细胞增殖和分化都有深远影响。已知许多人类细胞系对RA的生长抑制作用敏感。我们分析了已建立的源自人类实体瘤的细胞系对RA的敏感性。通过[3H]胸苷掺入和细胞增殖检测RA在单层培养中对细胞生长的抑制作用。在此我们报告,11种广泛使用的人类细胞系对RA耐药。大多数源自癌(A - 431、BT - 20、C - 41、ACHN、HCT116、293、A549和PA - 1);两种源自肉瘤(Saos - 2和A673);一种是黑色素瘤细胞系(A - 375)。由于核类视黄醇受体与RA的生物学效应有关,我们通过Northern印迹法和针对RARβ的核糖核酸酶保护分析,检测了RA耐药细胞系中维甲酸受体(RARs)RARα、RARβ、RARγ以及类视黄醇X受体(RXRs)RXRα、RXRβ和RXRγ的表达。RARα转录本在所有细胞系中均组成性表达。相比之下,RARβ仅在7种RA耐药细胞系(Saos - 2、ACHN、293、A549、A - 375、A673和PA - 1)中表达,并且在某些情况下其水平会被RA增强。在大多数细胞系中,RARγ表达较低且不受RA影响。RXR基因在所选细胞系组中表现出非常独特的表达模式。一般来说,RXRα是表达最丰富的亚型,RXRβ表达水平较低,且无法检测到RXRγ。在任何RA耐药细胞系中,RXR表达均不受RA调节。此处呈现的结果表明,这些人类肿瘤细胞系对RA的耐药性不能简单地与RAR或RXR或两者的表达相关联。