SEPARATION OF PARATHYROID HORMONE-LIKE ACTIVITY FROM TRANSFORMING GROWTH FACTOR-ALPHA AND FACTOR-BETA IN THE CANINE ADENOCARCINOMA (CAC-8) MODEL OF HUMORAL HYPERCALCEMIA OF MALIGNANCY
SEPARATION OF PARATHYROID HORMONE-LIKE ACTIVITY FROM TRANSFORMING GROWTH FACTOR-ALPHA AND FACTOR-BETA IN THE CANINE ADENOCARCINOMA (CAC-8) MODEL OF HUMORAL HYPERCALCEMIA OF MALIGNANCY
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DOI：
10.1210/endo-124-5-2456
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发表时间：
1989-05-01
期刊：
影响因子：
4.8
通讯作者：
CAPEN, CC
中科院分区：
文献类型：
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作者：
MERRYMAN, JI;ROSOL, TJ;CAPEN, CC
评价恶性体液性高钙血症的犬腺癌模型（CAC-8）的转化生长因子（TGF）-α。和-β，PTH样活性[腺苷酸环化酶刺激活性（阿克萨）]和体外骨吸收活性。通过反相或阳离子交换HPLC分离CAC-8中存在的生物活性。TGF.alpha.从TGF β中分离肿瘤提取物。和阿克萨。TGF.alpha.在26-30%乙腈之间洗脱，并通过RIA进行鉴定。在初始的反相分离后，TGF β。和阿克萨在36-38%乙腈之间共洗脱。依次进行阳离子交换，然后进行反相HPLC分离TGF β。来自阿克萨。使用体外骨吸收试验评价含有阿克萨的组分，证明了阿克萨的共纯化和骨吸收活性。PTH受体拮抗剂[Nle 8，18，Tyr 34]牛PTH-（3-34）-酰胺，但不是[Nle 8，18，Tyr 34]牛PTH-（7-34）-酰胺，完全抑制柱洗脱液中的阿克萨。来自CAC-8原代培养物的条件细胞培养基主要含有潜伏的TGF β。可以被酸化激活。这些发现表明癌症相关高钙血症的CAC-8模型产生PTH样因子TGF α，和TGF β。其可通过反相或阳离子交换HPLC分离。这一特征有助于探讨TGF和PTH样蛋白在恶性肿瘤体液性高钙血症发病机制中的作用。
A canine adenocarcinoma model (CAC-8) of humoral hypercalcemia of malignancy was evaluated for transforming growth factors (TGF)-.alpha. and -.beta., PTH-like activity [adenylate cyclase-stimulating activity (ACSA)], and in vitro bone-resorbing activity. Biological activities present in CAC-8 were separated by reverse phase or cation exchange HPLC. TGF.alpha. in tumor extract was separated from TGF.beta. and ACSA by reverse phase HPLC. TGF.alpha. eluted between 26-30% acetonitrile and was identified by RIA. After the initial reverse phase separation, TGF.beta. and ACSA in tumor extract coeluted between 36-38% acetonitrile. Sequential cation exchange followed by reverse phase HPLC separated TGF.beta. from ACSA. Evaluation of fractions containing ACSA using an in vitro bone-resorbing assay demonstrated copurification of ACSA and bone-resorbing activity. The PTH receptor antagonist [Nle8,18,Tyr34]bovine PTH-(3-34)-amide, but not [Nle8,18,Tyr34]bovine PTH-(7-34)-amide, completely inhibited ACSA in column eluates. Conditioned cell culture medium from CAC-8 primary cultures contained predominantly latent TGF.beta. that could be activated by acidification. These findings indicate that the CAC-8 model of cancer-associated hypercalcemia produces a PTH-like factor, TGF.alpha., and TGF.beta. that were separable by reverse phase or cation exchange HPLC. This feature should be useful to investigate the role of TGFs and PTH-like proteins in the pathogenesis of humoral hypercalcemia of malignancy.