Real-time imaging analysis of androgen receptor function in hormone refractory Prostate cancer

激素难治性前列腺癌雄激素受体功能的实时成像分析

• 批准号: 14571514
• 负责人: UKIMURA Osamu
• 金额: $ 2.5万
• 依托单位: Kyoto Prefectural University of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571514/
• 关键词: Prostate cancer; androgen receptor; green fluorescent protein; hormone refractory Prostate cancer; green fluorescent protein

Androgen has been shown to promote cell proliferation of prostate cancer through the action of androgen receptor (AIR) that is a number of the nuclear receptor superfamily. In a prostate cancer cell line, LNCaP, a point mutation (T877A) is found in AR gene, and it has been supposed that this mutation would be involved in hypersensitivity for low concentration of androgen. In the present study, trafficking of AR and AR(T877A) in living prostate and non-prostate cancer cell lines under various concentration of the ligands was examined by tagging green fluorescent protein (GFP) to the receptors. In a non-prostate cancer cell line, COS-1,wild type AR and mutant AR were differentially localized in the absence of ligand ; AR was completely localized in cytoplasm, but AR(T877A) was localized in cytoplasm and nucleus. In contrast, both AR and AR(T877A) were localized in cytoplasm and nucleus in LNCaP. Upon the addition of the ligand, both type of the receptors that localized in the cytoplasm were translocated into the nucleus in the both cell lines. Moreover, in the presence of low concentration of androgen, the translocation was observed in LNCaP cells, but not in COS-1 cells, and there was no significant difference in the translocation pattern between wild type AR and mutant AR. These results suggest that hypersensitivity of AR may be affected by intracellular factors, such as co-factors and crosstalk with another signal pathway, more than the mutation of AR gene. Real-time imaging with GFP was a promising tool for the investigation of molecular mechanism of hormone refractory appearance prostate cancer.

雄激素通过其核受体超家族中的雄激素受体（AIR）促进前列腺癌细胞增殖。在前列腺癌细胞系LNCaP中，发现AR基因的一个点突变（T877 A），并推测该突变可能与对低浓度雄激素的超敏反应有关。在本研究中，贩运AR和AR（T877 A）在活的前列腺和非前列腺癌细胞系在不同浓度的配体检查标记绿色荧光蛋白（GFP）的受体。在非前列腺癌细胞系中，COS-1、野生型AR和突变型AR在不存在配体的情况下差异定位; AR完全定位于细胞质中，但AR（T877 A）定位于细胞质和细胞核中。而在LNCaP中，AR和AR（T877 A）均定位于细胞质和细胞核。在添加配体后，定位于细胞质中的两种类型的受体在两种细胞系中均移位到细胞核中。此外，在低浓度雄激素存在下，LNCaP细胞中可观察到易位，而COS-1细胞中未观察到易位，且野生型AR和突变型AR之间的易位模式无显著差异。这些结果表明，AR的超敏反应可能受到细胞内因素的影响，如辅因子和与另一条信号通路的串扰，而不是AR基因的突变。GFP实时成像技术是研究激素抵抗性前列腺癌发生机制的有效工具。