Department of Biochemistry and Molecular Biology, Poznan University of Medical Sciences, Święcickiego 6 Street, 60-781 Poznan, Poland.
Department of Thoracic Surgery, Poznan University of Medical Sciences, Szamarzewskiego 62 Street, 60-569 Poznan, Poland.
Lung Cancer. 2015 Feb;87(2):107-16. doi: 10.1016/j.lungcan.2014.12.012. Epub 2014 Dec 25.
Recent studies indicated that estrogens may influence the development of non-small cell lung cancer (NSCLC). The 17-beta-hydroxysteroid dehydrogenase type 1 (HSD17B1) catalyzes the reduction of estrone (E1) to the highly potent E2. Although the significance of aromatase in an intratumoral E2 production in NSCLC is well established, the role of HSD17B1 remains largely unknown. Therefore, we investigated the expression of HSD17B1 in lung cancerous and corresponding histopathologically unchanged tissues from NSCLC patients and the association between HSD17B1 expression and clinicopathological features. Than, we examined the biological significance of HSD17B1 in NSCLC cells in vitro. We tested the impact of 5-Aza-2'-deoxycytidine (5-dAzaC) on HSD17B1 expression and activity.
We used Real Time quantitative PCR (RT-qPCR), Western blotting and immunohistochemistry to evaluate HSD17B1 expression in tissues obtained from 48 patients with NSCLC. The methylation status of the promoter region of HSD17B1 in A549 and Calu-1 cells was evaluated by bisulfite sequencing. We investigated the effect of 5-dAzaC on HSD17B1 transcript levels (by RT-qPCR) and on HSD17B1 enzyme activity by measuring the conversion of E1 to E2. The xCELLigence System was used for monitoring of cell proliferation.
We found a substantial increase of HSD17B1 mRNA and protein amount in NSCLC tissues compared with histopathologically unchanged tissues in the group of male patients. An overexpression of HSD17B1 was associated with squamous cell carcinoma and with lung cancer stage 3A. We showed that 5-dAzaC induces DNA demethylation of HSD17B1 promoter, leading to increased HSD17B1 mRNA levels and protein activity in NSCLC cells. It resulted in enhanced E2 production in both cell lines and supported the proliferation of Calu-1 cells but not A549 cells.
Increased expression of HSD17B1 in NSCLC may contribute to an elevated intratissue level of E2 and consequently may support the development and spread of cancer.
最近的研究表明，雌激素可能会影响非小细胞肺癌（NSCLC）的发展。17-β-羟类固醇脱氢酶 1（HSD17B1）催化将雌酮（E1）还原为高活性的雌二醇（E2）。尽管芳香酶在 NSCLC 中肿瘤内产生 E2 中的意义已得到充分证实，但 HSD17B1 的作用在很大程度上仍不清楚。因此，我们研究了 HSD17B1 在 NSCLC 患者的肺癌组织和相应的组织学未改变组织中的表达，以及 HSD17B1 表达与临床病理特征之间的关系。然后，我们在体外检查了 HSD17B1 在 NSCLC 细胞中的生物学意义。我们检测了 5-氮杂-2'-脱氧胞苷（5-dAzaC）对 HSD17B1 表达和活性的影响。
我们使用实时定量 PCR（RT-qPCR）、Western 印迹和免疫组织化学评估了 48 名 NSCLC 患者组织中 HSD17B1 的表达。通过亚硫酸氢盐测序评估 A549 和 Calu-1 细胞中 HSD17B1 启动子区域的甲基化状态。我们通过 RT-qPCR 研究了 5-dAzaC 对 HSD17B1 转录水平的影响，并通过测量 E1 向 E2 的转化来研究 HSD17B1 酶活性。xCELLigence 系统用于监测细胞增殖。
与男性患者组中组织学未改变的组织相比，我们发现 NSCLC 组织中 HSD17B1 的 mRNA 和蛋白含量大量增加。HSD17B1 的过表达与鳞状细胞癌和肺癌 3A 期有关。我们表明，5-dAzaC 诱导 HSD17B1 启动子的 DNA 去甲基化，导致 NSCLC 细胞中 HSD17B1 mRNA 水平和蛋白活性增加。这导致在两种细胞系中 E2 的产生增加，并支持 Calu-1 细胞的增殖，但不支持 A549 细胞的增殖。
NSCLC 中 HSD17B1 的表达增加可能导致组织内 E2 水平升高，从而可能支持癌症的发展和扩散。
