Fibulin-1 is Down-Regulated Through Promoter Hypermethylation and Suppresses Renal Cell Carcinoma Progression
Fibulin-1 is Down-Regulated Through Promoter Hypermethylation and Suppresses Renal Cell Carcinoma Progression
复制标题
Fibulin-1 通过启动子高甲基化下调并抑制肾细胞癌进展
DOI：
10.1016/j.juro.2013.01.098
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发表时间：
2013-07-01
影响因子：
6.6
通讯作者：
Xu, Hua
中科院分区：
文献类型：
--
作者：
Xiao, Wei;Wang, Ji;Xu, Hua
目的：肾细胞癌是世界上最常见的癌症之一，但其分子机制尚未完全了解。Fibulin-1是一种多功能的细胞外基质蛋白，与多种癌症有关，但其在肾细胞癌中的功能尚不清楚。我们研究了fibulin-1在肾细胞癌中的表达和功能。材料和方法：我们采用实时聚合酶链反应、Western印迹分析和免疫组化方法来检测肾细胞癌细胞和患者组织中fibulin-1的表达。采用甲基化特异性聚合酶链反应（PCR）和定量测序技术检测FBLN 1基因启动子甲基化状态。结果：Fibulin-1在肾癌细胞系和患者组织中表达明显下调，在ACHN和786-O肾癌细胞中的表达明显下调。失调与肾细胞癌进展相关。FBLN 1基因启动子区高甲基化，其甲基化状态与fibulin-1表达相关。在体外培养的肾癌细胞和裸鼠移植瘤中，Fibulin-1的过表达显著降低了细胞的生长，增强了肿瘤细胞的凋亡，降低了细胞的运动性和血管生成。在肾细胞癌中作为肿瘤抑制因子和血管生成抑制因子发挥作用。
Purpose: Renal cell carcinoma is one of the most common cancers worldwide but the molecular mechanisms that underlie it are not fully understood. Fibulin-1, a multi-functional extracellular matrix protein, is involved in many types of cancer but its function in renal cell carcinoma is unclear. We investigated fibulin-1 expression and function in renal cell carcinoma.Materials and Methods: We performed real-time polymerase chain reaction, Western blot analysis and immunohistochemistry to determine fibulin-1 expression in renal cell carcinoma cells and patient tissues. Methylation specific polymerase chain reaction and quantitative sequencing were done to examine the methylation status of the FBLN1 gene promoter. Eukaryotic expression plasmid and a lentiviral vector were used to over express fibulin-1 in ACHN and 786-O renal cell carcinoma cells to investigate its function in vitro and in vivo.Results: Fibulin-1 was significantly down-regulated in renal cell carcinoma cell lines and patient tissues. Dysregulation was associated with renal cell carcinoma progression. The FBLN1 gene promoter region was hypermethylated and its methylation status correlated with fibulin-1 expression. Fibulin-1 over expression led dramatically to decreased cell growth, enhanced tumor cell apoptosis, decreased cell motility and angiogenesis in cultured renal cell carcinoma cells and in xenograft tumors in nude mice.Conclusions: Fibulin-1 is down-regulated in renal cell carcinoma through promoter hypermethylation. It functions as a tumor suppressor and angiogenesis inhibitor in renal cell carcinoma.