DIFFERENTIAL-EFFECTS OF GLUCOCORTICOID ON EXPRESSION OF SURFACTANT PROTEINS IN A HUMAN-LUNG ADENOCARCINOMA CELL-LINE
DIFFERENTIAL-EFFECTS OF GLUCOCORTICOID ON EXPRESSION OF SURFACTANT PROTEINS IN A HUMAN-LUNG ADENOCARCINOMA CELL-LINE
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DOI：
10.1016/0167-4889(88)90179-6
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发表时间：
1988-06-30
期刊：
影响因子：
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通讯作者：
WHITSETT, JA
中科院分区：
文献类型：
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作者：
OREILLY, MA;GAZDAR, AF;WHITSETT, JA
已在源自人肺腺癌的连续细胞系中鉴定出 Mr 28 000-36 000 (SP-A) 和 Mr 42 000-46 000 (proSP-B) 肺表面活性剂相关糖蛋白的合成。通过免疫印迹分析、ELISA测定和细胞的[35S]蛋氨酸标记来检测SP-A。 SP-A 作为内切糖苷酶 F 敏感糖蛋白分泌到培养基中，与通过 2D-IEF-SDS-PAGE 在人灌洗液中鉴定出的 SP-A 亚型共迁移。 细胞 RNA 与 SP-A 特异性 cDNA 的杂交鉴定出丰富的 2.2 kb mRNA 种类，与在人肺中观察到的相同。地塞米松以剂量依赖性方式显着抑制 SP-A RNA 和蛋白质含量。在相同的培养条件下，糖皮质激素显着刺激了一种独特的表面活性蛋白 SP-B 的合成。 SP-B 前体以异质 Mr 42 000-46 000 蛋白的形式分泌到培养基中，pI 4.6-5.1，并且对糖苷内切酶 F 敏感。糖皮质激素以剂量依赖性方式增强 proSP-B 的合成，并且与 Northern 印迹分析检测到的 2.0 kb SP-B mRNA 增加相关。该细胞系分泌proSP-B作为Mr 42 000-46 000糖基化蛋白，并且不加工Mr 7000-8000表面活性剂肽的前体。总之，已鉴定出合成和分泌两种表面活性剂相关蛋白 SP-A 和 proSP-B 的人腺癌细胞系。 糖皮质激素增强该细胞系中 SP-B 的表达，但抑制 SP-A 的表达。分泌表面活性蛋白的连续细胞系的鉴定可用于研究这些重要蛋白质的合成和分泌以及用于临床用途的蛋白质的生产。
Synthesis of pulmonary surfactant-associated glycoproteins of Mr 28 000-36 000 (SP-A) and Mr 42 000-46 000 (proSP-B) has been identified in a continuous cell line derived from a human lung adenocarcinoma. SP-A was detected by immunoblot analysis, ELISA assay and by [35S]methionine labelling of the cells. SP-A was secreted into the media as an endoglycosidase F sensitive glycoprotein which co-migrated with the isoforms of SP-A identified in human lavage fluid by 2D-IEF-SDS-PAGE. Hybridization of cellular RNA with SP-A specific cDNA identified an abundant 2.2 kb mRNA species, identical to that observed in human lung. SP-A RNA and protein content were markedly inhibited by dexamethasone in a dose-dependent fashion. Under identical culture conditions, synthesis of a distinct surfactant protein, SP-B, was markedly stimulated by the glucocorticoid. The SP-B precursor was secreted into the media as heterogenous Mr 42 000-46 000 protein, pI 4.6-5.1, and was sensitive to endoglycosidase F. Synthesis of proSP-B was enhanced by the glucocorticoid in a dose-dependent fashion and was associated with increased SP-B mRNA of 2.0 kb detected by Northern blot analysis. The cell line secreted proSP-B as Mr 42 000-46 000 glycosylated protein and did not process the precursor to the Mr 7000-8000 surfactant peptide. In summary, a human adenocarcinoma cell line has been identified which synthesizes and secretes two surfactant-associated proteins, SP-A and proSP-B. Glucocorticoid enhanced SP-B but inhibited SP-A expression in this cell line. The identification of a continuous cell line secreting surfactant proteins may be useful in the study of synthesis and secretion of these important proteins and for production of the proteins for clinical uses.