Pre-clinical Research Center, Tokyo Medical University, Tokyo, Japan.
Department of Molecular Pathology, Tokyo Medical University, Tokyo, Japan.
Oncogene. 2021 May;40(21):3695-3706. doi: 10.1038/s41388-021-01787-5. Epub 2021 May 4.
Since cervical cancer still afflicts women around the world, it is necessary to understand the underlying mechanism of cervical cancer development. Infection with HPV is essential for the development of cervical intraepithelial neoplasia (CIN). In addition, estrogen receptor signaling is implicated in the development of cervical cancer. Previously, we have isolated human wings apart-like (WAPL), which is expected to cause chromosomal instability in the process of HPV-infected precancerous lesions to cervical cancer. However, the role of WAPL in the development of CIN is still unknown. In this study, in order to elucidate the role of WAPL in the early lesion, we established WAPL overexpressing mice (WAPL Tg mice) and HPV E6/E7 knock-in (KI) mice. WAPL Tg mice developed CIN lesion without HPV E6/E7. Interestingly, in WAPL Tg mice estrogen receptor 1 (ESR1) showed reduction as compared with the wild type, but cell growth factors MYC and Cyclin D1 controlled by ESR1 expressed at high levels. These results suggested that WAPL facilitates sensitivity of ESR1 mediated by some kind of molecule, and as a result, affects the expression of MYC and Cyclin D1 in cervical cancer cells. To detect such molecules, we performed microarray analysis of the uterine cervix in WAPL Tg mice, and focused MACROD1, a co-activator of ESR1. MACROD1 expression was increased in WAPL Tg mice compared with the wild type. In addition, knockdown of WAPL induced the downregulation of MACROD1, MYC, and Cyclin D1 but not ESR1 expression. Furthermore, ESR1 sensitivity assay showed lower activity in WAPL or MACROD1 downregulated cells than control cells. These data suggested that WAPL increases ESR1 sensitivity by activating MACROD1, and induces the expression of MYC and Cyclin D1. Therefore, we concluded that WAPL not only induces chromosomal instability in cervical cancer tumorigenesis, but also plays a key role in activating estrogen receptor signaling in early tumorigenesis.
由于宫颈癌仍然困扰着世界各地的女性，因此有必要了解宫颈癌发展的潜在机制。HPV 感染是宫颈癌前病变发展为宫颈上皮内瘤变（CIN）的必要条件。此外，雌激素受体信号转导与宫颈癌的发生有关。以前，我们已经分离出人类 wings apart-like（WAPL），预计它会在 HPV 感染的癌前病变向宫颈癌发展的过程中引起染色体不稳定。然而，WAPL 在 CIN 发展中的作用仍不清楚。在这项研究中，为了阐明 WAPL 在早期病变中的作用，我们建立了 WAPL 过表达小鼠（WAPL Tg 小鼠）和 HPV E6/E7 敲入（KI）小鼠。WAPL Tg 小鼠在没有 HPV E6/E7 的情况下发展为 CIN 病变。有趣的是，与野生型相比，WAPL Tg 小鼠中的雌激素受体 1（ESR1）表达减少，但由 ESR1 控制的细胞生长因子 MYC 和细胞周期蛋白 D1 表达水平较高。这些结果表明，WAPL 通过某种分子促进 ESR1 介导的敏感性，从而影响宫颈癌细胞中 MYC 和细胞周期蛋白 D1 的表达。为了检测这种分子，我们对 WAPL Tg 小鼠的子宫颈进行了微阵列分析，并将雌激素受体 1 的共激活剂 MACROD1 作为研究重点。与野生型相比，WAPL Tg 小鼠中的 MACROD1 表达增加。此外，敲低 WAPL 诱导 MACROD1、MYC 和细胞周期蛋白 D1 的下调，但不影响 ESR1 的表达。此外，ESR1 敏感性测定显示，下调 WAPL 或 MACROD1 的细胞的活性低于对照细胞。这些数据表明，WAPL 通过激活 MACROD1 增加 ESR1 敏感性，并诱导 MYC 和细胞周期蛋白 D1 的表达。因此，我们得出结论，WAPL 不仅在宫颈癌肿瘤发生中诱导染色体不稳定，而且在早期肿瘤发生中还发挥激活雌激素受体信号转导的关键作用。
