Expression and functional significance of HtrA1 loss in endometrial cancer.
Expression and functional significance of HtrA1 loss in endometrial cancer.
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DOI：
10.1158/1078-0432.ccr-09-3069
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发表时间：
2011-02-01
期刊：
影响因子：
--
通讯作者：
Shridhar V
中科院分区：
文献类型：
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作者：
Mullany SA;Moslemi-Kebria M;Rattan R;Khurana A;Clayton A;Ota T;Mariani A;Podratz KC;Chien J;Shridhar V
本研究的目的是确定子宫内膜癌中丝氨酸蛋白酶HtrA1的缺失是否会促进EC细胞系的侵袭潜能。用Western印迹分析和免疫组织化学方法分别检测HtrA1在EC细胞系和原发肿瘤中的表达。通过迁移实验、侵袭实验和体内移植实验，比较HtrA1表达克隆和HtrA-1基因敲除克隆的转移情况。13个EC细胞系HtrA1的Western印迹分析显示，HtrA1在所有7个乳头状浆液性EC细胞系中的表达均完全丧失。在Hec1A和Hec1B细胞系中，HtrA1的下调导致侵袭潜能增加3-4倍。外源性HtrA1在Ark 1和Ark 2细胞中的表达导致这些细胞的侵袭和迁移能力降低3-4倍。与内源性HtrA1表达的对照细胞相比，Hec1B细胞中HtrA1下调相关的肺转移率增加。HtrA1在Ark 2细胞中的表达增强导致转移到肺部的肿瘤结节明显少于亲本或蛋白水解酶缺陷(SA突变)的Ark 2细胞。免疫组织化学(IHC)分析显示，57%(105/184)的原发EC肿瘤HtrA1低表达。HtrA1低表达与高级别子宫内膜样瘤的相关性有统计学意义(p=0.016)。总体而言，这些数据表明HtrA1的缺失可能与子宫内膜肿瘤的侵袭性和转移能力有关。
The purpose of this study was to determine if loss of serine protease HtrA1 in endometrial cancer will promote the invasive potential of EC cell lines. Western blot analysis and immunohistochemistry methods were used to determine HtrA1 expression in EC cell lines and primary tumors, respectively. Migration, invasion assays and in vivo xenograft experiment were performed to compare the extent of metastasis between HtrA1 expressing and HtrA-1 knocked down clones. Western blot analysis of HtrA1 in 13 EC cell lines revealed complete loss of HtrA1 expression in all 7 papillary serous EC cell lines. Downregulation of HtrA1 in Hec1A and Hec1B cell lines resulted in a 3-4 fold increase in the invasive potential. Exogenous expression of HtrA1 in Ark 1 and Ark 2 cells resulted in 3-4 fold decrease in both invasive and migration potential of these cells. There was an increased rate of metastasis to the lungs associated with HtrA1 downregulation in Hec1B cells compared to control cells with endogenous HtrA1 expression. Enhanced expression of HtrA1 in Ark 2 cells resulted in significantly less tumor nodules metastasizing to the lungs compared to parental or protease deficient (SA mutant) Ark 2 cells. Immunohistochemical (IHC) analysis showed 57% (105/184) of primary EC tumors had low HtrA1 expression. The association of low HtrA1 expression with high-grade endometrioid tumors was statistically significant (p=0.016). Collectively, these data indicate loss of HtrA1 may contribute to the aggressiveness and metastatic ability of endometrial tumors.