Effects of growth factors, hormones, bacterial lipopolysaccharides, and lipotechoic acids on the clonal growth of normal ureteral epithelial cells in serum-free culture.
Effects of growth factors, hormones, bacterial lipopolysaccharides, and lipotechoic acids on the clonal growth of normal ureteral epithelial cells in serum-free culture.
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生长因子、激素、细菌脂多糖和脂磷壁酸对无血清培养中正常输尿管上皮细胞克隆生长的影响。
DOI：
10.1002/jcp.1041500108
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发表时间：
1992
影响因子：
5.6
通讯作者：
Elgavish,A
中科院分区：
文献类型：
--
作者：
Wille,JJ;Park,J;Elgavish,A
采用体外组织培养技术研究细菌内毒素对正常人输尿管上皮细胞生长的影响。NHU细胞的原代培养物起始于人输尿管组织的外植体生长培养物，并在含1%胎牛血清（FCS）的F-12* 培养基中的胶原凝胶上培养。在含有牛垂体提取物（0.5% BPE）和0.05% BSA的F-12* 培养基中，以相对较低的接种细胞密度直接接种在塑料培养皿上的NHU细胞传代培养物实现了最佳克隆生长。结果表明，F-12* 培养基中的胰岛素可以被低三个数量级的IGF-1替代。进一步的克隆生长实验表明，PGE 1具有生长刺激作用，可以取代BPE作为生长因子的必需品。这一发现与BPE生长需要可以被霍乱毒素或二丁酰cAMP替代的事实一致。这些结果表明，BPE和霍乱毒素都是通过激活cAMP依赖性促有丝分裂途径来发挥作用的。测试了七种革兰氏阴性细菌脂多糖（LPS）和三种革兰氏阳性细菌脂儿茶酸（LT）对NHU克隆生长的影响。大肠杆菌055：B5、0128：B12和0127：B8菌株、肺炎克雷伯菌和铜绿假单胞菌的LPS在5 ~ 25 μg/ml的最低有效剂量范围内均表现出明显的生长抑制作用。LPS来源弗罗姆.大肠杆菌（0111：B4）在最高试验浓度（100 μg/ml）下无生长效应。而来源于化脓性链球菌、S.在1 μg/ml < [LT] < 50 μg/ml浓度范围内，粪肠球菌、金黄色葡萄球菌和枯草杆菌均显著促进克隆生长。LT来自链球菌。100 μg/ml的化脓性链球菌对克隆生长有抑制作用。LPS的生长抑制作用显示对生长培养基中氢化可的松的存在敏感，表明LPS对生长的影响是通过花生四烯酸级联介导的。我们推测，这些结果表明尿路上皮组织对尿路疾病中观察到的致病菌群的易感性与具有增殖能力的尿路上皮细胞对革兰氏阴性菌产生的LPS生长抑制的不同敏感性之间存在联系。然而，进一步的研究与尿路致病性血清型将是必要的，以证实这种可能性。革兰氏阳性菌产生的LT的生长刺激活性可能是由于其结合细胞相关纤连蛋白并激活纤连蛋白受体的能力，纤连蛋白受体是配体受体诱导的促有丝分裂跨膜信号传导途径的一部分。
In vitro tissue culture techniques were employed to study the effects of bacterial endotoxins on the growth of normal epithelial cells from the human ureter (NHU). Primary cultures of NHU cells were initiated from explant outgrowth cultures of human ureteral tissue and cultured on collagen gel in F‐12* medium containing 1% fetal calf serum (FCS). Optimal clonal growth of secondary cultures of NHU cells seeded at relatively low seeding cell densities, directly on plastic dishes, was achieved in F‐12* medium containing bovine pituitary extract (0.5% BPE) and 0.05% BSA. Results indicated that insulin in the F‐12* medium could be replaced by three orders of magnitude less IGF‐1. Further clonal growth experiments demonstrated that PGE1is growth stimulatory and can replace BPE as a growth factor requirement. This finding was in agreement with the fact that BPE growth requirement could be replaced by cholera toxin or dibutyryl cAMP. These results suggested that both BPE and cholera toxin operated by activation of a cAMP‐dependent mitogenic pathway. Seven gram‐negative bacterial lipopolysaccharides (LPS) and three gram‐positive bacterial lipotechoic acids (LT) were tested for their effects on NHU clonal growth. Three out of the five LPS derived fromEscherichia coli(strains 055:B5, 0128:B12, and 0127:B8), LPS fromKlebsiella pneumoniae, and LPS fromPseudomonas aeruginosaall showed significant growth inhibitory effects at minimally effective doses ranging from 5 to 25 μg/ml. LPS derived fromE. colistrain (0111:B4) had no growth effects at the highest concentration tested (100 μg/ml). In contrast, LT derived fromStreptococcus pyogenes, S. faecalis, Staphylococcus aureas, andBacillus subtilisall markedly enhanced clonal growth at concentrations ranging from 1 μg/ml < [LT] < 50 μg/ml. LT fromStrep. pyogeneswas inhibitory to clonal growth at 100 μg/ml. The growth inhibitory effects of LPS were shown to be sensitive to the presence of hydrocortisone in the growth medium, indicating that LPS effects on growth are mediated via the arachidonic acid cascade. We speculate that these results indicate a link between the susceptibility of uroepithelial tissue to the pathogenic microflora seen in urinary tract diseases and the differential sensitivity of proliferation‐competent uroepithelial cells to growth inhibition by LPS produced by gram‐negative bacteria. However, further studies with uropathogenic serotypes will be necessary to corroborate this possibility. The growth‐stimulating activity of LTs produced by gram‐positive bacteria may be due to their ability to bind to cell‐associated fibronectin and to activate the fibronectin receptor as part of ligand receptor–induced mitogenic transmembrane signalling pathway.