Establishment and characterization of androgen-independent human prostate cancer cell lines, LN-REC4 and LNCaP-SF, from LNCaP
Establishment and characterization of androgen-independent human prostate cancer cell lines, LN-REC4 and LNCaP-SF, from LNCaP
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DOI：
10.1111/j.1442-2042.2007.01532.x
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发表时间：
2007-03-01
影响因子：
2.6
通讯作者：
Namiki, Mikio
中科院分区：
文献类型：
--
作者：
Iwasa, Yoichi;Mizokami, Atsushi;Namiki, Mikio
目的：为了研究前列腺癌 (PCa) 中雄激素非依赖性生长的机制，我们从雄激素依赖性 PCa 细胞系 LNCaP 中建立了两种 PCa 细胞系 LN-REC4 和 LNCaP-SF。 材料和方法：LN-Pre 和 LN-REC4 细胞分别从完整和去势的严重联合免疫缺陷 (SCID) 小鼠上生长的 LNCaP 肿瘤中产生。在无类固醇条件下体外培养LNCaP细胞6个月后，建立了LNCaP-SF细胞。为了显示LN-REC4和LNCaP-SF细胞的特征，通过检查生长速率来研究雄激素敏感性，并通过逆转录聚合酶链式反应（RT-PCR）检查前列腺特异性抗原（PSA）、雄激素受体（AR）、p21、p27和细胞周期蛋白D1的表达。使用条件培养基进行体外血管生成测定。为了检测血管内皮生长因子（VEGF）的表达水平，还进行了RT-PCR和酶联免疫吸附测定。结果和结论：在去势小鼠中，LN-REC4细胞比LNCaP细胞增殖更好，并且与去势无关，尽管LN-REC4细胞对雄激素的反应性在体外减弱程度小于LNCaP细胞，但无论是否去势，LN-REC4细胞都表现良好。去势小鼠中的 LNCaP-SF 细胞增殖速度比正常小鼠更快。 LNCaP-SF细胞中PSA的表达仍然受到雄激素的诱导。 LN-REC4 和 LNCaP-SF 细胞中 AR、p21、p27 和 cyclin D1 的表达没有变化。血管生成测定显示两种细胞均刺激血管生成。 LN-REC4 比 LNCaP 和 LN-Pre 细胞更能诱导 VEGF。然而，LNCaP-SF 中每个细胞的 VEGF 表达低于 LNCaP 细胞，表明其他因素可能参与血管生成。这些细胞系可能是研究雄激素非依赖性生长和复发性前列腺癌治疗的有用工具。
Aim: To investigate the mechanisms of androgen-independent growth in prostate cancer (PCa), we established two PCa cell lines, LN-REC4 and LNCaP-SF, from the androgen-dependent PCa cell line, LNCaP.Materials and methods: LN-Pre and LN-REC4 cells were generated from LNCaP tumors grown on intact and castrated severe combined immunodeficient (SCID) mouse, respectively. After we cultured LNCaP cells under a steroid-free conditions for 6 months in vitro, LNCaP-SF cells were established. To show the character of LN-REC4 and LNCaP-SF cells, androgen sensitivity was investigated through examination of growth rate, and prostate-specific antigen (PSA), androgen receptor (AR), p21, p27, and cyclin D1 expression were examined by reverse transcription-polymerase chain reaction (RT-PCR). Angiogenesis assay in vitro was carried out using conditioned medium. To examine the expression level of vascular endothelial growth factor (VEGF), RT-PCR and enzyme-linked immunosorbent assay were also done.Results and conclusions: LN-REC4 cells proliferated better than LNCaP cells in castrated mice and did well irrespective of castration, although responsiveness for androgen of LN-REC4 cells attenuated less than that of LNCaP cells in vitro. LNCaP-SF cells in castrated mice proliferated more rapidly than in normal mice. The PSA expression in LNCaP-SF cells was still induced by androgen. Expression of AR, p21, p27 and cyclin D1 were not changed in LN-REC4 and LNCaP-SF cells. Angiogenesis assay showed that both cells stimulated angiogenesis. LN-REC4 induced VEGF more than LNCaP and LN-Pre cells. However, expression of VEGF per cell in LNCaP-SF was lower than LNCaP cells, suggesting that other factors might be involved in angiogenesis. These cell lines might be a useful tool for researching androgen-independent growth and treatments of recurred PCa.