Zhang Liansuo, Shao Wei, Li Mingqing, Liu Songping
Department of Obstetrics and Gynecology, Jinshan Hospital, Fudan University, Shanghai 201508, China.
Laboratory for Reproductive Immunology, Hospital of Obstetrics and Gynecology, Fudan University, Shanghai 200080, China.
Biomedicines. 2023 Sep 11;11(9):2506. doi: 10.3390/biomedicines11092506.
Post-translational modification of proteins is involved in the occurrence of endometriosis (EM); however, the role of ubiquitination modification in EM remains unclear. Integrin β3 (ITGB3) is one of the β-subunits of integrins, which plays a key role in tumor progression. In this study, we investigated the roles of ITGB3 and ITCH, one of the ubiquitin E3 ligases, in ectopic endometrial stromal cells (ESCs) and EM. Primary ectopic ESCs and normal ESCs were isolated and purified. Western blot was used to detect the expression of ITGB3 and ITCH in ESCs. The interaction between ITGB3 and ITCH in ESCs was investigated by the co-immunoprecipitation and ubiquitylation analysis. With or without the overexpression of ITCH and/or ITGB3, the proliferation and invasion of ectopic ESCs were detected by the CCK8 assay and transwell migration assay, respectively. We found that ITGB3 is upregulated in ectopic ESCs from patients with EM. ITCH interacts with ITGB3 by co-immunoprecipitation, and ITCH-overexpressing significantly increased the ubiquitination of ITGB3. The data of the CCK8 assays showed that ITGB3 overexpression significantly promoted cell proliferation of ectopic ESCs at 12, 24, 48, and 72 h. The transwell migration assays showed that ITGB3 overexpression significantly enhanced the invasive ability. However, ITCH had the opposite effects in both assays. Our findings indicate that ITCH-mediated ubiquitylation of ITGB3 regulates the proliferation and invasion ability of ectopic ESCs in EM.
蛋白质的翻译后修饰参与了子宫内膜异位症（EM）的发生；然而，泛素化修饰在EM中的作用仍不清楚。整合素β3（ITGB3）是整合素的β亚基之一，在肿瘤进展中起关键作用。在本研究中，我们探讨了ITGB3和泛素E3连接酶之一ITCH在异位子宫内膜基质细胞（ESC）和EM中的作用。分离并纯化了原发性异位ESC和正常ESC。采用蛋白质免疫印迹法检测ESC中ITGB3和ITCH的表达。通过免疫共沉淀和泛素化分析研究了ESC中ITGB3与ITCH之间的相互作用。分别通过CCK8法和Transwell迁移试验检测了ITCH和/或ITGB3过表达与否时异位ESC的增殖和侵袭能力。我们发现，EM患者异位ESC中ITGB3表达上调。ITCH通过免疫共沉淀与ITGB3相互作用，过表达ITCH显著增加了ITGB3的泛素化。CCK8试验数据显示，ITGB3过表达在12、24、48和72小时显著促进异位ESC的细胞增殖。Transwell迁移试验表明，ITGB3过表达显著增强了侵袭能力。然而，ITCH在这两种试验中均有相反的作用。我们的研究结果表明，ITCH介导的ITGB3泛素化调节了EM中异位ESC的增殖和侵袭能力。
