BIS-(BETA-CHLOROETHYL)SULFIDE (BCES)-INDUCED CHANGES IN EPIDERMAL-CELL HOMEOSTASIS INVITRO
BIS-(BETA-CHLOROETHYL)SULFIDE (BCES)-INDUCED CHANGES IN EPIDERMAL-CELL HOMEOSTASIS INVITRO
复制标题
DOI：
10.1016/0041-008x(88)90358-4
复制
发表时间：
1988-09-30
影响因子：
3.8
通讯作者：
BERNSTEIN, IA
中科院分区：
文献类型：
--
作者：
KU, WW;BERNSTEIN, IA
建立了大鼠皮肤角质形成细胞培养体系，研究了发泡剂双(β-氯乙基)硫(BCES)对细胞增殖和分化动态平衡的影响。凝集素用于揭示角质形成细胞分化过程中细胞表面碳水化合物的变化。在新生大鼠的表皮中，异凝素，单叶格里芬I-B4(GS I-B4)结合在基底层细胞表面。Ulex uropeus凝集素I(UEA)结合在棘状细胞和下层颗粒细胞表面，因此被认为是角质形成细胞分化的指标。建立了一种测定低钙培养大鼠皮肤角质形成细胞原代单层培养中结合的UEA与结合的GS I-B4的比率(UEA B4比率)的荧光分析方法。UEA B4比率被发现分别代表了单层培养中分化和增殖细胞室的相对大小(W.W.Ku和I.A.Bernstein，1988，Exp.Cell Res.，175,298-316)。与溶剂对照组相比，在第1天暴露于BCES 1小时的单层培养在第7天显示出与剂量相关的UEA B4比率的增加。凝集素结合位点的分析结果表明，BCES暴露导致GS I-B4结合减少，而UEA结合几乎没有变化，这是导致UEA B4比率增加的原因之一。放射自显影显示，暴露于BCES的单层细胞在复制DNA合成过程中也表现出早期的扰动。在复制DNA合成的干扰之后，暴露于BCES的培养物不能通过有丝分裂将细胞产生成单层。除了UEA B4比率的增加外，BCES暴露的单层细胞也表现出与剂量相关的DNA丢失，并在第7天出现扩大的细胞。这些扩大的细胞没有显示出DNA合成的证据，这些细胞组显示出强烈的UEA染色，只有微弱的GS I-B4染色。总体而言，暴露于低浓度的BCES似乎破坏了单层培养系统中细胞增殖和分化的正常动态平衡。这种破坏主要是由于萌发(基底)细胞比例的减少，伴随而来的是一些早期分化的细胞，可能是早期的棘状细胞或棘状细胞。
A rat cutaneous keratinocyte culture system was developed to study the effects of the vesicant bis-(β-chloroethyl)sulfice (BCES) on the homeostasis of cell proliferation and differentiation. Lectins were used to reveal cell surface carbohydrate changes as the keratinocytes differentiate. In the newborn rat epidermis, the isolectin, Griffonia simplicifolia I-B4(GS I-B4), binds to basal cell surfaces. Ulex europeus agglutinin I (UEA) binds to the surfaces of spinous and lower granular cells and is therefore considered an indicator of keratinocyte differentiation. A fluorometric assay was developed which determines the ratio of bound UEA to bound GS I-B4(the UEA B4ratio) in primary monolayer cultures of rat cutaneous keratinocytes maintained in low Ca2+medium. The UEA B4ratio was found to be a representation of the relative sizes of the differentiating and proliferating cell compartments in the monolayer cultures, respectively (W. W. Ku and I. A. Bernstein, 1988, Exp. Cell Res., 175, 298–316). Monolayer cultures exposed for 1 hr to BCES at Day 1 exhibited a dose-related increase in the UEA B4ratio at Day 7 when compared to solvent controls. The results from the analysis of lectin binding sites showed a decrease in GS I-B4binding with little or no change in UEA binding as a result of BCES exposure, contributing to the increase in the UEA B4ratio. BCES-exposed monolayers also showed early perturbations in replicative DNA synthesis as revealed by autoradiography. Subsequent to the perturbations in replicative DNA synthesis was an inability of BCES-exposed cultures to produce cells into the monolayer through mitosis. In addition to an increase in the UEA B4ratio, BCES-exposed monolayers also showed a dose-related loss of DNA, with the appearance of enlarged cells at Day 7. These enlarged cells failed to show evidence of DNA synthesis, with groups of these cells showing intense UEA staining with only faint GS I-B4staining. Overall, exposure to low concentrations of BCES appeared to disrupt the normal homeostasis of cell proliferation and differentiation in this monolayer culture system. This disruption was primarily through a reduction in the fraction of germinative (basal) cells with concomitant retention of some early differentiated cells, presumably early spinous or spinous cells.