New chemotherapeutic drug sensitivity assay for colon carcinomas in monolayer culture.
New chemotherapeutic drug sensitivity assay for colon carcinomas in monolayer culture.
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发表时间：
1988-06
期刊：
影响因子：
11.2
通讯作者：
P. Schroy;A. Cohen;S. Winawer;E. Friedman
中科院分区：
文献类型：
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作者：
P. Schroy;A. Cohen;S. Winawer;E. Friedman
在原代单层培养物中测试了十种先前未经治疗的结肠癌的化疗药物敏感性。结肠癌被部分消化成上皮细胞群，这些上皮细胞群以 42 +/- 9% (SE) 的平均效率铺在无血清培养基中的 I 型胶原蛋白-牛血清白蛋白底物上，产生紧密贴壁的上皮细胞斑块。通过细胞角蛋白、上皮细胞表面表位、连接复合物和刷状缘的存在来判断培养的细胞为上皮细胞。将每个癌组织置于 40 至 60 个培养皿 (35 mm) 中，每个培养皿平均产生 28 +/- 8 (SE) 个集落（6832 +/- 1952 个细胞）。在双份板中测试的药物为丝裂霉素 C、顺铂、链脲佐菌素和 5-氟尿嘧啶，浓度为 0.1、1、10 和 100 微克/ml，以及血清中峰值耐受药物浓度的 0.1、1 和 2 倍。铺板后24小时，除去任何非贴壁细胞，并将贴壁肿瘤细胞连续暴露于药物3天。每种药物在反应性肿瘤中以剂量依赖性方式诱导集落溶解。通过[3H]胸苷掺入未通过药物处理裂解的集落中来鉴定耐药的循环细胞。十种癌症中的每一种都表现出对临床可达到的浓度范围内的一种或多种化疗药物的固有耐药性。
Ten previously untreated colon carcinomas were tested for chemotherapeutic drug sensitivity in primary monolayer culture. Colon carcinomas were partly digested to groups of epithelial cells which plated with a mean efficiency of 42 +/- 9% (SE) on a collagen I-bovine serum albumin substrate in serum-free medium, producing patches of tightly adherent epithelial cells. The cultured cells were judged epithelial by the presence of cytokeratins, an epithelial cell surface epitope, junctional complexes, and brush borders. Each carcinoma was plated in 40 to 60 Petri dishes (35 mm), yielding a mean of 28 +/- 8 (SE) colonies per dish (6832 +/- 1952 cells). Drugs tested in duplicate plates were mitomycin C, cisplatin, streptozotocin, and 5-fluorouracil at 0.1, 1, 10, and 100 micrograms/ml, and at 0.1, 1, and 2x the peak tolerated drug concentration in serum. Twenty-four h after plating, any nonadherent cells were removed, and the adherent tumor cells were continuously exposed to the drugs for 3 days. Each drug induced colony lysis in a dose-dependent manner in responsive tumors. Drug-resistant, cycling cells were identified by [3H]thymidine incorporation in colonies which were not lysed by drug treatment. Each of the ten carcinomas exhibited inherent resistance to one or more chemotherapy drugs within the concentration ranges clinically achievable.