Stable differentiation of a human colon adenocarcinoma cell line by sodium butyrate is associated with multidrug resistance.
Stable differentiation of a human colon adenocarcinoma cell line by sodium butyrate is associated with multidrug resistance.
复制标题
丁酸钠对人结肠腺癌细胞系的稳定分化与多药耐药性相关。
DOI：
10.1002/jcp.1041600202
复制
发表时间：
1994
影响因子：
5.6
通讯作者：
Kim,YS
中科院分区：
文献类型：
--
作者：
Ho,SB;Yan,PS;Dahiya,R;Neuschwander-Tetri,BA;Basbaum,C;Kim,YS
结直肠癌通常由代表不同分化细胞系的细胞类型组成，然而，关于这些癌症的分化与耐药之间的关系却知之甚少。本研究旨在建立和鉴定稳定的人结肠癌细胞系LS174T的分化克隆，并研究该细胞系与其未分化亲本细胞系之间的耐药性。将LS174T细胞用0.5 mM的丁酸钠诱导30天后，在标准培养液中进行再培养。出现扁平细胞灶，用克隆环分离，并进行亚克隆。其中一个亚克隆命名为LS174T-D。在无丁酸钠的标准培养条件下，LS174T-D克隆保持了稳定的分化表型。它的特征是形成一个带有圆顶的极化单层，并存在突出的顶端微绒毛和紧密连接。与亲本细胞系相比，该细胞系在软琼脂和裸鼠体内的生长受到抑制。LS174T-D细胞表达免疫反应阳性的肠粘蛋白抗原和刷状缘酶二肽氨基肽酶(DAP-IV)和氨基肽酶。LS174T-D细胞的DAP-IV和氨基肽酶活性分别是亲本细胞的5.6倍和3.4倍。增殖实验显示，与亲代细胞株相比，LS174T-D细胞对阿霉素(93倍)、顺铂(23倍)、5-氟尿嘧啶(12倍)、5-氟脱氧尿苷(31倍)和甲氨蝶呤(12.5倍)的耐药性更强。分化和未分化细胞系对(~3H)-5-氟代脱氧尿苷的摄取无显著差异。Western印迹和RNA Northern印迹检测两种细胞株的MDR-1 P-糖蛋白水平也同样较低。然而，LS174T-D细胞的总谷胱甘肽含量和谷胱甘肽-S转移酶活性分别是亲本细胞的6倍和3倍。用DL-丁硫氨酸亚磺胺处理谷胱甘肽，可逆转LS174T-D对顺铂的耐药性。丁酸钠长期治疗可诱导或选择具有肠细胞分化特征的结肠癌细胞。这种稳定分化的细胞系与谷胱甘肽介导的多药耐药相关，并为进一步研究正常和癌变的分化提供了一个模型。©1994 Wiley-Liss，Inc.
Colorectal cancers are often composed of cell types representing various differentiated cell lineages, however little is known concerning the relationship of differentiation and drug resistance in these cancers. The present study was performed to develop and characterize a stable, differentiated clone of the human colon cancer cell line LS174T and to characterize the drug resistance of this cell line in relation to its undifferentiated parental cell line. LS174T cell line was treated with the differentiating agent sodium butyrate (0.5 mM) for 30 days, then recultured in standard medium. Foci of flat‐appearing cells appeared and were isolated using cloning rings, and subcloned. One subclone was designated LS174T‐D. The LS174T‐D clone maintains a stable, differentiated phenotype in standard culture conditions in the absence of sodium butyrate. It is characterized by the formation of a polarized monolayer with dome formation and the presence of prominent apical microvilli and tight junctions. This cell line demonstrated reduced growth in soft agar and nude mice compared with the parental cell line. LS174T‐D cells expressed immunoreactive intestinal mucin antigens and brush border enzymes dipeptidyl aminopeptidase (DAP)‐IV and aminopeptidase. The activities of DAP‐IV and aminopeptidase were increased 5.6‐fold and 3.4‐fold, respectively, in LS174T‐D compared with parental cells. Proliferation assays demonstrated that, compared with the parental cell line, LS174T‐D cells were more resistant to doxorubicin (93‐fold), cisplatin (23‐fold), 5‐fluorouracil (12‐fold), 5‐fluorodeoxyuridine (31‐fold), and methotrexate (12.5‐fold). Intracellular uptake of (3H)‐5‐fluorodeoxyuridine did not differ significantly in the differentiated and undifferentiated cell lines. Levels of mdr‐1 p‐glycoprotein measured by Western blot and RNA Northern blot assays were also similarly low in both cell lines. However, total glutathione content and glutathione‐S‐transferase activities were increased in LS174T‐D cells by sixfold and threefold, respectively, compared with parental cells. Depletion of glutathione by pretreatment with DL‐buthionine sulfoximine reversed LS174T‐D resistance to cisplatin. Long‐term treatment with sodium butyrate induces or selects for colon cancer cells with features of enterocytic differentiation. This stably differentiated cell line is associated with glutathione‐mediated multidrug resistance, and provides a model for further studies of differentiation in normal and cancerous colon. © 1994 Wiley‐Liss, Inc.