WATANABE Manabu, CHEN Iris Yun-Ju, ISHIKURA Yasuhiro, NAKAGAWA Takayuki, MOCHIZUKI Manabu, NISHIMURA Ryohei, SASAKI Nobuo, SUGANO Sumio
Laboratory of Functional Genomics, Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Minato-ku, Tokyo, Japan.
J Vet Med Sci. 2009 Nov;71(11):1413-7. doi: 10.1292/jvms.001413.
In this study, we examined the effects of four chemical reagents, 5 azacytidine (5azaC), all-trans-retinoic acid (ATRA), Phorbol 12-myristate 13-acetate (TPA) and trichostatin A (TSA), on an MCM-B2 canine mammary tumor cell line. Growth of the MCM-B2 cells was inhibited by addition of any of these four chemical reagents in a dose-dependent manner. TPA-treated cells became blast-cell-like and had high expressions of cyclin D, cyclin B and PCNA. Both ATRA-treated and 5azaC-treated cells showed decreased expression of cell cycle related molecules and increased expressions of the mammary epithelial marker cytokeratin 18 and underwent morphological changes. ATRA-treated cells were converted from the originally spindle-shaped cells into cubic-shaped cells, and 5azaC-treated cells became fibroblast-like. In the TSA-treated cells, the cytoplasm was enlarged, increasing the cytoplasm/nucleus ratio, and the expressions of cell cycle-promoting molecules (cyclin D and PCNA), as well as cell cycle-inhibiting factors (p21(WAF1) and p27(kip1)), were increased. These results demonstrated the growth-inhibiting, differentiation-inducing and antitumor effects of each of these four chemicals on the unique phenotype of canine mammary tumors.
在本研究中，我们检测了四种化学试剂，即5-氮杂胞苷（5azaC）、全反式维甲酸（ATRA）、佛波酯12-肉豆蔻酸酯13-乙酸酯（TPA）和曲古抑菌素A（TSA），对MCM-B2犬乳腺肿瘤细胞系的影响。添加这四种化学试剂中的任何一种，MCM-B2细胞的生长均受到剂量依赖性抑制。经TPA处理的细胞变成母细胞样，且细胞周期蛋白D、细胞周期蛋白B和增殖细胞核抗原（PCNA）表达较高。经ATRA处理和5azaC处理的细胞均显示细胞周期相关分子表达降低，乳腺上皮标志物细胞角蛋白18表达增加，并发生形态学改变。经ATRA处理的细胞从最初的纺锤形细胞转变为立方形细胞，经5azaC处理的细胞变成成纤维细胞样。在经TSA处理的细胞中，细胞质增大，细胞质/细胞核比值增加，促进细胞周期的分子（细胞周期蛋白D和PCNA）以及抑制细胞周期的因子（p21（WAF1）和p27（kip1））的表达均增加。这些结果证明了这四种化学物质对犬乳腺肿瘤独特表型的生长抑制、诱导分化和抗肿瘤作用。
