Xinxiang Key Laboratory of Tumor Migration, Invasion and Precision Medicine, Henan Key Laboratory of Immunology and Targeted Drugs, School of Laboratory Medicine, Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, Xinxiang Medical University, Xinxiang, Henan Province, People's Republic of China.
Department of Oncology, Dong Ying People' S Hospital, Dongying, Shandong Province, People's Republic of China.
Cell Commun Signal. 2022 Apr 7;20(1):45. doi: 10.1186/s12964-022-00861-z.
Breast cancer is the most common cancer in women worldwide. More than 70% of breast cancers are estrogen receptor (ER) alpha positive. Compared with ER alpha-negative breast cancer, which is more aggressive and has a shorter survival time, ER alpha-positive breast cancer could benefit from endocrine therapy. Selective estrogen receptor modulators, such as tamoxifen, are widely used in endocrine therapy. Approximately half of ER alpha-positive breast cancer patients will eventually develop endocrine resistance, making it a major clinical challenge in therapy. Thus, decoding the throughput of estrogen signaling, including the control of ER alpha expression and stability, is critical for the improvement of breast cancer therapeutics.
TRIM3 and ER alpha protein expression levels were measured by western blotting, while the mRNA levels of ER alpha target genes were measured by RT-PCR. A CCK-8 assay was used to measure cell viability. RNA sequencing data were analyzed by Ingenuity Pathway Analysis. Identification of ER alpha signaling activity was accomplished with luciferase assays, RT-PCR and western blotting. Protein stability assays and ubiquitin assays were used to detect ER alpha protein degradation. Ubiquitin-based immunoprecipitation assays were used to detect the specific ubiquitination modification on the ER alpha protein.
In our current study, we found that TRIM3, an E3 ligase, can promote ER alpha signaling activity and breast cancer progression. TRIM3 depletion inhibits breast cancer cell proliferation and migration, while unbiased RNA sequencing data indicated that TRIM3 is required for the activity of estrogen signaling on the -genome-wide scale. The immunoprecipitation assays indicated that TRIM3 associates with ER alpha and promotes its stability, possibly by inducing K63-linked polyubiquitination of ER alpha. In conclusion, our data implicate a nongenomic mechanism by which TRIM3 stabilizes the ER alpha protein to control ER alpha target gene expression linked to breast cancer progression.
Our study provides a novel posttranslational mechanism in estrogen signaling. Modulation of TRIM3 expression or function could be an interesting approach for breast cancer treatment. Video abstract.
乳腺癌是全世界女性最常见的癌症。超过 70%的乳腺癌雌激素受体（ER）α阳性。与更具侵袭性、生存时间更短的 ERα阴性乳腺癌相比，ERα阳性乳腺癌可以受益于内分泌治疗。选择性雌激素受体调节剂，如他莫昔芬，广泛用于内分泌治疗。大约一半的 ERα阳性乳腺癌患者最终会发展为内分泌抵抗，这是治疗中的一个主要临床挑战。因此，解码雌激素信号通路的通量，包括 ERα表达和稳定性的控制，对于改善乳腺癌治疗至关重要。
通过 Western blot 测定 TRIM3 和 ERα 蛋白表达水平，通过 RT-PCR 测定 ERα 靶基因的 mRNA 水平。用 CCK-8 测定细胞活力。用 Ingenuity Pathway Analysis 分析 RNA 测序数据。用荧光素酶测定、RT-PCR 和 Western blot 测定 ERα 信号活性。用蛋白稳定性测定和泛素测定检测 ERα 蛋白降解。用泛素免疫沉淀测定检测 ERα 蛋白上的特异性泛素化修饰。
在我们目前的研究中，我们发现 E3 连接酶 TRIM3 可以促进 ERα 信号活性和乳腺癌的进展。TRIM3 耗竭抑制乳腺癌细胞增殖和迁移，而无偏 RNA 测序数据表明，TRIM3 在全基因组范围内对雌激素信号的活性是必需的。免疫沉淀测定表明，TRIM3 与 ERα 结合并促进其稳定性，可能通过诱导 ERα 的 K63 连接多泛素化。总之，我们的数据提示了一种非基因组机制，通过该机制，TRIM3 稳定 ERα 蛋白以控制与乳腺癌进展相关的 ERα 靶基因表达。
我们的研究提供了雌激素信号的一种新的翻译后机制。调节 TRIM3 的表达或功能可能是治疗乳腺癌的一种有趣方法。视频摘要。
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