A microplate assay for selective measurement of growth of epithelial tumor cells in direct coculture with stromal cells.
A microplate assay for selective measurement of growth of epithelial tumor cells in direct coculture with stromal cells.
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发表时间：
2004-05
影响因子：
2
通讯作者：
M. Kawada;Y. Yoshimoto;K. Minamiguchi;H. Kumagai;T. Someno;Tohru Masuda;M. Ishizuka;D. Ikeda
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文献类型：
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作者：
M. Kawada;Y. Yoshimoto;K. Minamiguchi;H. Kumagai;T. Someno;Tohru Masuda;M. Ishizuka;D. Ikeda
基质细胞通过扩散因子和粘附在调节上皮恶性肿瘤中发挥重要作用。肿瘤-基质细胞相互作用的调节是新的抗肿瘤策略的一个有吸引力的靶点。为了筛选相互作用的调节剂，我们现在开发了一种定量比色法，用于测量肿瘤细胞与基质细胞共培养中使用罗丹尼尔蓝染料的生长情况。Rhodanile蓝在共培养中特异性染色细胞角蛋白阳性肿瘤细胞。将人前列腺癌细胞LNCaP、PC-3和DU-145与正常前列腺基质细胞（PrSC）在微孔板中共培养，用罗丹尼尔蓝染色法选择性地测定共培养中前列腺癌细胞的生长情况。利用这个系统，我们在临床使用的药物和天然产物之间寻找肿瘤-基质细胞相互作用的调节剂。结果，我们发现5-氟尿嘧啶、博来霉素和苯恶唑啉a与PrSC共培养对前列腺癌细胞的抑制作用比单独培养更强。无需预先标记细胞和转染标记基因，我们的新方法简单，快速，因此可用于筛选肿瘤-基质细胞相互作用的调节剂。此外，我们的研究结果表明，低分子量化合物调节肿瘤-基质细胞相互作用。
Stromal cells play an important role in regulating epithelial malignancies through diffusible factors and adhesion. Modulation of the tumor-stromal cell interaction is an attractive target for new antitumor strategies. To screen for a modulator of the interaction, we have now developed a quantitative colorimetric assay for measurement of tumor cell growth in coculture with stromal cells using rhodanile blue dye. Rhodanile blue specifically stained cytokeratin-positive tumor cells in the coculture. When human prostate carcinoma cells LNCaP, PC-3 and DU-145 were cocultured with normal prostate stromal cells (PrSC) in a microplate, growth of the prostate cancer cells in the coculture was selectively measured by the rhodanile blue staining method. Using this system, we searched for a modulator of the tumor-stromal cell interaction among clinically used drugs and natural products. As a result, we found that 5-fluorouracil, bleomycin and phthoxazolin A inhibit prostate cancer cell growth more strongly in coculture with PrSC than that in monoculture. Without need to pre-label cells and transfect a marker gene, our new method is simple, rapid and thus useful for screening for modulators of the tumor-stromal cell interaction. Furthermore, our results suggest that low molecular weight compounds modulate the tumor-stromal cell interaction.