Shabana A H, Lecolle S, Goldberg M, Forest N
Laboratoire de Biologie-Odontologie, Université Paris 7, Institut Biomédical des Cordeliers, France.
Microsc Res Tech. 1998 Nov 1;43(3):233-41. doi: 10.1002/(SICI)1097-0029(19981101)43:3<233::AID-JEMT5>3.0.CO;2-L.
Protein kinase inhibitor H-7 was reported to stimulate desmosome formation in normal keratinocytes and to inhibit proliferation of neural cell lines. In the present study, the effects of this inhibitor on adhesion and growth of KB human oral carcinoma cells were investigated. H-7 was found to enhance desmosome assembly, as evidenced by an increased punctate labeling for the major desmosomal markers. Immunogold labeling confirmed the formation of desmosomes both at the cell surface and in the cytoplasm. In order to assess cell proliferation and possible correlation with adhesion, confluent cultures were treated and both adherert and detached cell fractions were counted. Under serum-free conditions, H-7 significantly reduced cell detachment. In contrast, EGF stimulated cell detachment, and this effect was abolished when cells were simultaneously treated with both EGF and H-7. Total cell counts were also significantly reduced by H-7, both in the presence and absence of EGF. Using the TUNEL technique, labeled cells were increased after H-7 treatment, thus implicating protein kinase inhibition in cell death. These results indicate that H-7 inhibits growth and stimulates adhesion of KB carcinoma cells.
据报道，蛋白激酶抑制剂H-7可刺激正常角质形成细胞中的桥粒形成，并抑制神经细胞系的增殖。在本研究中，研究了该抑制剂对KB人口腔癌细胞黏附和生长的影响。发现H-7可增强桥粒组装，主要桥粒标记物的点状标记增加证明了这一点。免疫金标记证实了细胞表面和细胞质中桥粒的形成。为了评估细胞增殖以及与黏附的可能相关性，对汇合培养物进行处理，并对贴壁和 detached 细胞部分进行计数。在无血清条件下，H-7显著减少细胞脱离。相反，表皮生长因子（EGF）刺激细胞脱离，当细胞同时用EGF和H-7处理时，这种作用被消除。无论有无EGF，H-7均显著减少细胞总数。使用末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）技术，H-7处理后标记细胞增加，因此表明蛋白激酶抑制与细胞死亡有关。这些结果表明，H-7抑制KB癌细胞的生长并刺激其黏附。
