HK-2 - AN IMMORTALIZED PROXIMAL TUBULE EPITHELIAL-CELL LINE FROM NORMAL ADULT HUMAN KIDNEY
HK-2 - AN IMMORTALIZED PROXIMAL TUBULE EPITHELIAL-CELL LINE FROM NORMAL ADULT HUMAN KIDNEY
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DOI：
10.1038/ki.1994.6
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发表时间：
1994-01-01
影响因子：
19.6
通讯作者：
TOROKSTORB, B
中科院分区：
文献类型：
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作者：
RYAN, MJ;JOHNSON, G;TOROKSTORB, B
评估近端肾小管细胞 (PTC) 生理学和病理生理学机制的研究越来越多地利用细胞培养系统来避免整个器官/整个动物实验的复杂性。然而，目前不存在源自成人肾脏的分化良好的 PTC 系。因此，本研究的目标是通过转导人乳头瘤病毒（HPV 16）E6/E7 基因来建立这样的系。将来自正常成人肾皮质的原代 PTC 培养物暴露于含有 HPV 16 E6/E7 基因的重组逆转录病毒，产生称为 HK-2（人肾 2）的细胞系，该细胞系在无血清培养基中连续生长一年多。 HK-2 细胞的生长依赖于表皮生长因子，并且细胞保留了分化良好的 PTC 的表型（碱性磷酸酶、γ 谷氨酰转肽酶、亮氨酸氨肽酶、酸性磷酸酶、细胞角蛋白、α3β1 整联蛋白、纤连蛋白呈阳性；因子 VIII 相关抗原、6.19 抗原和 CALLA 内肽酶呈阴性）。此外，HK-2细胞保留近端肾小管上皮的功能特征（Na+依赖性/根皮苷敏感性糖转运；腺苷酸环化酶对甲状旁腺的反应性，但对抗利尿剂、激素没有反应）。通过 PCR 测定，E6/E7 基因存在于 HK-2 基因组中。为了评估其作为研究损伤和修复工具的潜在用途，将 HK-2 细胞暴露于有毒浓度的 H2O2 +/- 铁螯合剂（去铁胺）或羟自由基清除剂（苯甲酸钠）疗法。只有前者阻断了 H2O2 细胞毒性，重现了之前用新鲜分离的大鼠近端肾小管段获得的结果。总之，通过转导 HPV 16 E6/E7 基因建立了永生化成人 PTC 系。根据其组织化学、免疫细胞化学和功能特征，它似乎分化良好，并且可以重现用新鲜分离的 PTC 获得的实验结果。因此，这种新的 PTC 系列可能具有大量的研究应用。
Studies assessing mechanisms of proximal tubular cell (PTC) physiology and pathophysiology increasingly utilize cell culture systems to avoid the complexity of whole organ/whole animal experiments. However, no well-differentiated PTC line derived from adult human kidney currently exists. Therefore, the goal of this research was to establish such a line by transduction with human papilloma virus (HPV 16) E6/E7 genes. A primary PTC culture from normal adult human renal cortex was exposed to a recombinant retrovirus containing the HPV 16 E6/E7 genes, resulting in a cell line designated HK-2 (human kidney-2) which has grown continuously in serum free media for more than one year. HK-2 cell growth is epidermal growth factor dependent and the cells retain a phenotype indicative of well-differentiated PTCs (positive for alkaline phosphatase, gamma glutamyltranspeptidase, leucine aminopeptidase, acid phosphatase, cytokeratin, alpha3beta1 integrin, fibronectin; negative for factor VIII-related antigen, 6.19 antigen and CALLA endopeptidase). Furthermore, HK-2 cells retain functional characteristics of proximal tubular epithelium (Na+ dependent/phlorizin sensitive sugar transport; adenylate cyclase responsiveness to parathyroid, but not to antidiuretic, hormone). The E6/E7 genes are present in the HK-2 genome, as determined by PCR. To assess its potential usefulness as a tool for studying injury and repair, HK-2 cells were exposed to a toxic concentration of H2O2 +/- iron chelation (deferoxamine) or hydroxyl radical scavenger (Na benzoate) therapy. Only the former blocked H2O2 cytotoxicity, reproducing results previously obtained with freshly isolated rat proximal tubular segments. In conclusion, an immortalized adult human PTC line has been established by transduction with HPV 16 E6/E7 genes. It appears to be well-differentiated on the basis of its histochemical, immune cytochemical, and functional characteristics, and it can reproduce experimental results obtained with freshly isolated PTCs. Thus, this new PTC line could have substantial research application.