Low STING expression promotes endometrial stromal cell invasion and migration via the STING/IRF-3/IFN-β1 pathway in eutopic endometrium of women with endometriosis

Abstract

Aims: The primary aim of the current study was to elucidate the function of the stimulator of interferon genes (STING) in the eutopic endometrium of women with endometriosis. Materials and Methods: STING expression and signaling pathways were verified by western blot analysis and immunohistochemistry after si-STING treatment. Cell proliferation and invasion and migration were assessed using 5-ethynyl-2’-deoxyuridine and transwell assays, respectively. Results: Within endometriosis tissues, STING was primarily expressed in the stroma of the eutopic endometrium and glandular epithelium of the ectopic endometrium. However, STING expression was significantly lower in the eutopic endometrium of patients with endometriosis compared to controls (p < 0.05). Additionally, cell proliferation (0.2866 ± 0.01470 vs. 0.6911 ± 0.01796, ^****p < 0.0001), invasion (130.0 ± 6.296 vs. 424.1 ± 22.31, ^****p < 0.0001), and migration (82.93 ± 6.940 vs. 82.93 ± 6.940, ^****p < 0.0001) were significantly increased in the si-STING groups. Moreover, following si-STING transfection, the expression of phosphorylated IRF-3 and TBK1 that are involved in STING/IRF3/IFNb1 signaling pathway decreased. The addition of exogenous IFN-β1 effectively increased stromal cell invasion (IFN-β1-NC vs. IFN-β1-si-STING 274.7 ± 7.767 vs. 135.7 ± 12.63, ***p < 0.0001) and migration (IFN-β1-NC and IFN-β1-si-STING 28.53 ± 3.625 vs. 28.53 ± 3.625, ***p < 0.0001) without significantly impacting cell proliferation (si-STING vs. IFN-1β-si-STING 0.6874 ± 0.02081 vs. 0.7187 ± 0.02638, p = 0.795). Conclusions: The STING signaling pathway plays an important role in endometrial stromal cell proliferation, invasion and migration associated with endometriosis.

摘要

目的

本研究的主要目的是阐明干扰素基因刺激因子(STING)在子宫内膜异位症女性正位子宫内膜的功能。

材料与方法

si-STING处理后, 通过免疫印迹分析和免疫组化验证STING的表达及信号通路。分别用5-乙基-2 ‘脱氧尿苷（EdU）和transwell法评估细胞增殖和侵袭迁移。

结果

在子宫内膜异位症组织中, STING主要表达于正位子宫内膜间质和异位子宫内膜腺上皮。但与对照组相比, 子宫内膜异位症患者正位子宫内膜STING表达明显降低(p < 0.05)。此外, 细胞增殖(0.2866±0.01470 vs 0.6911 ± 0.01796, ****p < 0.0001), 侵袭性(130.0±6.296 vs. 424.1±22.31, ****p < 0.0001)、迁移性(82.93±6.940 vs. 82.93±6.940, ****p < 0.0001) 在si-STING组显著增加。此外, si-STING转染后, 参与STING/iRF3/iFNb1信号通路的磷酸化iRF-3和tBK1的表达降低。外源iFN-β1的添加有效地增加了间质细胞的侵袭(iFN-β-nc vs. iFN-β1-si-STING 274.7±7.767 vs. 135.7±12.63, ***p<0.0001)和迁移(iFN-β1-nc和iFN-β1-si-STING 28.53±3.625 vs. 28.53±3.625, ***p<0.0001), 而对细胞增殖无显著影响(si-STING vs. iFN-1β-si-STING 0.6874±0.02081 vs. 0.7187±0.02638,p = 0.795)。

结论

STING信号通路在与子宫内膜异位症相关的子宫内膜间质细胞增殖、侵袭和迁移中起重要作用。

Keywords:

• Endometriosis
• eutopic endometrium
• cell invasion and migration
• innate immunity

关键词:

• 子宫内膜异位症
• 内膜
• 细胞侵袭和迁移
• 先天免疫

Acknowledgments

We thank Editage (www.editage.cn) for English language editing. We also thank all patients for their participation in this study.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This work was supported by the National Natural Science Foundation of China (grant number: 82071617).