TIPE2 inhibits the migration and invasion of endometrial cells by targeting β-catenin to reverse epithelial–mesenchymal transition

Abstract

STUDY QUESTION Do changes in tumor necrosis factor-α-induced protein 8 (TNFAIP8)-like 2 (TIPE2) levels in endometrium of patients with adenomyosis alter the proliferation, migration and invasion ability of endometrial cells? SUMMARY ANSWER TIPE2 expression levels were low in eutopic and ectopic endometrium of adenomyosis patients, and TIPE2 inhibited the migration and invasion of endometrial cells, mainly by targeting β-catenin, to reverse the epithelial-mesenchymal transition (EMT). WHAT IS KNOWN ALREADY Adenomyosis is a benign disease, but it has some pathophysiological characteristics similar to the malignant tumor. TIPE2 is a novel negative immune regulatory molecule, and it also participates in the development of malignant tumors. STUDY DESIGN, SIZE, DURATION Control endometrium (n = 48 women with non-endometrial diseases) and eutopic/ectopic endometrium from patients with adenomyosis (n = 50), human endometrial cancer cell lines, and primary endometrial cells from the eutopic endometrium of adenomyosis patients were used in the study. PARTICIPANTS/MATERIALS, SETTING, METHODS The expression level of TIPE2 mRNA and protein in the eutopic/ectopic endometrial tissues of adenomyosis patients and control endometrium was determined by quantitative RT-PCR (qRT-PCR), western blot and immunohistochemistry. The effects of TIPE2 overexpression and knockdown on the proliferation, migration and invasion of endometrial cell lines and primary adenomyotic endometrial cells were determined using a cell counting kit-8, 5-ethynyl-2′-deoxyuridine assay, colony-forming assay, transwell migration assay and matrigel invasion assay. The expression of EMT-related markers and signal molecules was detected by western blot. The interaction between TIPE2 and β-catenin was detected by co-immunoprecipitation and laser confocal microscopy. MAIN RESULTS AND THE ROLE OF CHANCE The mRNA and protein expression levels of TIPE2 in the eutopic and ectopic endometrial tissues of adenomyosis patients were significantly downregulated compared with the control endometrium (P ˂ 0.01). TIPE2 could bind to β-catenin and inhibit the nuclear translocation of β-catenin, downregulate the expression of stromal cell markers, upregulate the expression of glandular epithelial cell markers, decrease the occurrence of epithelial-mesenchymal transition (EMT) and suppress the migration and invasion of endometrial cells (P ˂ 0.01). LARGE SCALE DATA N/A. LIMITATIONS, REASONS FOR CAUTION In this study, the experiments were performed only in eutopic and ectopic endometrial tissues, endometrial cancer cell lines and primary adenomyotic endometrial cells. A mouse model of adenomyosis will be constructed to detect the effects of TIPE2 in vivo. WIDER IMPLICATIONS OF THE FINDINGS These results suggest that TIPE2 is involved in the development of adenomyosis, which provides a potential new diagnostic and therapeutic strategy for the treatment of adenomyosis. STUDY FUNDINGS/COMPETING INTEREST(S) This present study was supported by grants from the National Natural Science Foundation of China (81471437, 81771554), Natural Science Foundation of Shandong (ZR2018MH013), Science and technology development plan provided by Health and Family Planning Committee in Shandong (2014-25). The authors declare that they have no conflicts of interest.

Single-cell Transcriptomic Analysis of Eutopic Endometrium and Ectopic Lesions of Adenomyosis

“…In the follow-up comparison of the epithelial cell groups, rst because of the small number of epithelial cells in the AM_CTRL group and second because of the potential individual differences, only the AM_EC and AM_EM samples were compared, and the results show that the function of upregulated genes (p value < 0.05, fold change > 1.5) mainly focused on CMI-related terms. These results are consistent with the malignant tumour features of AM, including invasion and migration [21,29], and a subcluster1 with active motility is identi ed in epithelial cells. Thus, the present study support the theory of AM derived from the invasion and migration of endometrium.…”

“…3D). It has been reported that the pathogenesis of AM is closely related to the enhancement of invasion and migration of endometrial cells and epithelialmesenchymal transition (EMT) [29,30]; therefore, the cell motility of epithelial cell clusters became our focus. The DEGs of epithelial cells in the AM_EC versus AM_EM groups were analysed, and ITGA2, ITGB8 and CXCL8 were also identi ed in the upregulated genes with a log 2 fold change > 1 (Fig.…”

Tumor Necrosis Factor-α-Induced Protein 8-Like 2 Negatively Regulates Innate Immunity Against RNA Virus by Targeting RIG-I in Macrophages

“…Tumor necrosis factor-α-induced protein 8-like 2 is involved in the regulation of various physiological functions including inflammation, immunity, apoptosis, and cancer (11)(12)(13)(17)(18)(19)(20)(21). It has been reported that TIPE2 is a negative regulator required for maintaining immune homeostasis via inhibiting the activation of activator protein 1 (AP-1), NF-κB, and MAPK to negatively regulate the responses mediated by TCR and TLR (11,45,46).…”

“…TIPE2 deficiency in mice spontaneously develops fatal systemic inflammation and premature death, and loss of TIPE2 augments the production of pro-inflammatory cytokines upon stimulated with various inflammatory insults (16). Accelerating evidence indicates that TIPE2 inhibits the development of a variety of malignant tumors by suppressing cell proliferation (17,18), inducing cancer cell apoptosis (19), inhibiting tumor migration (20,21), and promoting antitumor immune responses mediated by CD8 + T and natural killer (NK) cells (22). TIPE2 has been recently shown to regulate the functional polarization of myeloid-derived suppressor cells (MDSCs) and maybe a potential therapeutic target for cancer immunotherapy (23).…”

<p>Regulatory Roles of Tumor Necrosis Factor-α-Induced Protein 8 Like-Protein 2 in Inflammation, Immunity and Cancers: A Review</p>

“…91,92 Moreover, it was defined that TIPE2 also induced CD8 + T cells and natural killer (NK) cells to secrete more cytokines, like interferon-γ (IFN-γ) and TNF-α, which in turn enhanced CD8 + T cells and NK cells' cytotoxicity and antitumor immune responses in spleen and tumor microenvironment, ultimately inhibiting the development and metastasis of breast cancer cells 93 (Table 1). In endometrial cancer cells, TIPE2 bound with β-catenin and decreased its nuclear translocation, suppressing EMT and tumorigenesis of endometrial cancer cells 94 (Table 1). The expression level of TIPE2 was extremely weak or undetectable in bladder, cervical and ovarian cancers, but the mechanisms remain unknown currently 34,35,95 (Table 1).…”