Kimball E S, Bohn W H, Cockley K D, Warren T C, Sherwin S A
Cancer Res. 1984 Aug;44(8):3613-9.
Reverse-phase high-performance liquid chromatography (HPLC) performed on urine from cancer patients and normal controls revealed the presence of seven chromatographically distinct peaks of transforming growth factor (TGF) activity, as measured by colony formation of normal rat kidney cells in soft agar. Comparison of urines from normal donors and cancer patients showed no differences in EGF (epidermal growth factor)-dependent beta-TGF-like activity but did reveal distinct patterns of EGF-related, EGF-independent alpha-TGF-like activity. All urine samples contained at least two chromatographically distinguishable forms of EGF-dependent TGF activity, eluting from HPLC as broad peaks with 30 and 43% acetonitrile. The remaining five TGFs eluted as sharp peaks with 32, 34, 35, 37, and 38% acetonitrile, demonstrated EGF-competing activity, and thus were functionally related to EGF. Two of the five EGF-related TGFs were consistently elevated only in the urine of cancer patients and eluted with 32% (TGFA) and 37% (TGFD) acetonitrile Two of the other EGF-related TGFs, eluting with 34% (TGFB) and 35% (TGFC) acetonitrile, were commonly found in both normals and cancer patients. The fifth EGF-related TGF, TGFE, eluting with 38% acetonitrile, was found only in normal donor specimens. TGFA corresponded to the unique Mr 30,000 TGF activity previously identified only in the urine of cancer patients. These observations demonstrate that cancer patients produce high levels of EGF-related TGF activities which can be readily distinguished, using reverse-phase HPLC, from EGF-related TGFs produced by normal individuals. Using a solid-phase competitive radioreceptor binding assay for EGF, we demonstrated that quantitation of EGF-competing activity is as sensitive and effective as the soft-agar colony formation assay for distinguishing HPLC profiles of urinary TGF from cancer patients versus that from normal individuals.
对癌症患者和正常对照者的尿液进行反相高效液相色谱（HPLC）分析，结果显示，通过正常大鼠肾细胞在软琼脂中的集落形成来测定，存在7个色谱上不同的转化生长因子（TGF）活性峰。正常供体和癌症患者尿液的比较表明，表皮生长因子（EGF）依赖性β-TGF样活性无差异，但确实揭示了与EGF相关的、不依赖EGF的α-TGF样活性的不同模式。所有尿液样本均含有至少两种色谱上可区分的EGF依赖性TGF活性形式，从HPLC中洗脱时为宽峰，乙腈含量分别为30%和43%。其余5种TGF以尖锐峰形式洗脱，乙腈含量分别为32%、34%、35%、37%和38%，表现出EGF竞争活性，因此在功能上与EGF相关。5种与EGF相关的TGF中有2种仅在癌症患者尿液中持续升高，分别以32%（TGFA）和37%（TGFD）乙腈洗脱。另外2种与EGF相关的TGF，分别以34%（TGFB）和35%（TGFC）乙腈洗脱，在正常人和癌症患者中均常见。第5种与EGF相关的TGF，TGFE，以38%乙腈洗脱，仅在正常供体样本中发现。TGFA对应于先前仅在癌症患者尿液中鉴定出的独特的分子量为30,000的TGF活性。这些观察结果表明，癌症患者产生高水平的与EGF相关的TGF活性，使用反相HPLC可以很容易地将其与正常个体产生的与EGF相关的TGF区分开来。使用针对EGF的固相竞争放射受体结合测定法，我们证明，对EGF竞争活性的定量与软琼脂集落形成测定法一样灵敏有效，可用于区分癌症患者与正常个体尿液TGF的HPLC谱。
