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锌指蛋白样蛋白 1 的 RING 结构域对子宫内膜癌细胞系 RL95-2 的增殖是必需的。

RING domain of zinc finger protein like 1 is essential for cell proliferation in endometrial cancer cell line RL95-2.

机构信息

Department of Clinical Laboratory, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China.

Department of Clinical Laboratory, Shandong University School hospital, Jinan, Shandong 250010, China.

出版信息

Gene. 2018 Nov 30;677:17-23. doi: 10.1016/j.gene.2018.07.053. Epub 2018 Jul 20.

Abstract

Endometrial cancer (EC) is the fourth most common cancer in women and exhibits increasing incidence and mortality. Some reports showed that the 5-year survival rate of EC was closely associated with the diagnosed stage. It is urgent to screen for sensitive and specific targets to improve early detection and EC therapy. In our study, we found that zinc finger protein like 1 (ZFPL1) was highly expressed in EC tissues and the EC cell line RL95-2, as detected via RT-qPCR and western blot analysis. Immunocytochemistry results showed that ZFPL1 was localized in the Golgi complex dependent on the C-terminal transmembrane domain. The MTT and EdU stains were employed to examine the effect of ZFPL1 on cell proliferation. We found that the silencing of ZFPL1 blocked cell proliferation and the expression of p-Akt308 and p-Akt473 but improved the protein level of PTEN. The overexpression of ZFPL1 and ZFPL1ΔTMD (deletion of the transmembrane domain) promoted cell proliferation and induced the expression of p-Akt308 and p-Akt473. However, the overexpression of ZFPL1ΔRING (deletion of the RING domain) caused loss of the function of ZFPL1 in cell proliferation and the PI3K/Akt pathway. In summary, ZFPL1 induced RL95-2 cell proliferation and was involved in PI3K/Akt pathway, suggesting the oncogenic role of ZFPL1 during EC development. Additionally, the RING domain was essential for the function of ZFPL1. These findings provided a new biomarker for EC diagnosis and therapy.

摘要

子宫内膜癌(EC)是女性中第四常见的癌症,发病率和死亡率呈上升趋势。一些报道表明,EC 的 5 年生存率与诊断阶段密切相关。迫切需要筛选出敏感和特异的靶点,以提高早期检测和 EC 治疗的效果。在我们的研究中,我们发现锌指蛋白样 1(ZFPL1)在 EC 组织和 EC 细胞系 RL95-2 中高表达,通过 RT-qPCR 和 Western blot 分析检测到。免疫细胞化学结果表明,ZFPL1 定位于高尔基复合体,依赖于 C 端跨膜结构域。MTT 和 EdU 染色用于检测 ZFPL1 对细胞增殖的影响。我们发现,ZFPL1 的沉默阻断了细胞增殖和 p-Akt308 和 p-Akt473 的表达,但提高了 PTEN 的蛋白水平。ZFPL1 和 ZFPL1ΔTMD(跨膜结构域缺失)的过表达促进了细胞增殖,并诱导了 p-Akt308 和 p-Akt473 的表达。然而,ZFPL1ΔRING(环指结构域缺失)的过表达导致 ZFPL1 在细胞增殖和 PI3K/Akt 通路中的功能丧失。总之,ZFPL1 诱导 RL95-2 细胞增殖,并参与 PI3K/Akt 通路,提示 ZFPL1 在 EC 发展过程中的致癌作用。此外,环指结构域对于 ZFPL1 的功能是必不可少的。这些发现为 EC 的诊断和治疗提供了一个新的生物标志物。

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