ZNF213 negatively controls triple negative breast cancer progression via Hippo/YAP signaling.

Breast cancer is one of the most commonly diagnosed malignancies worldwide, while the triple negative breast cancer (TNBC) is the most aggressive and virulent subtype in breast cancers. Compared with luminal type breast cancers, which could be well controlled by endocrine treatment, TNBC is worse in prognosis and lack of effective targeted therapy. Thus, it would be interesting and meaningful to identify novel therapeutic targets for TNBC treatments. Recent genomic data showed the activation of Hippo/YAP signaling in TNBC, indicating its critical roles in TNBC carcinogenesis and cancer progression. Hippo/YAP signaling could subject to several kinds of protein modifications, including ubiquitination and phosphorylation. Quite a few studies have demonstrated these modifications, which controlled YAP protein stability and turnover, played critical role in Hippo signaling activation In our current study, we identified ZNF213 as a negative modifier for Hippo/YAP axis. ZNF213 depletion promoted TNBC cell migration and invasion, which could be rescued by further YAP silencing. ZNF213 knocking down facilitated YAP protein stability and Hippo target gene expression, including CTGF and CYR61. Further mechanism studies demonstrated that ZNF213 associated with YAP and facilitated YAP K48-linked poly-ubiquitination at several YAP lysine sites (K252, K254, K321 and K497). Besides, the clinical data showed that ZNF213 negatively correlated with YAP protein level and Hippo target gene expression in TNBC samples. ZNF213 expression correlated with good prognosis in TNBC patients. Our data provided novel insights in YAP proteolytic regulation and TNBC progression.

ZNF213 Facilitates ER Alpha Signaling in Breast Cancer Cells.

BACKGROUND: Breast cancer is the most common women malignancy worldwide, while estrogen receptor alpha positive type accounts for two third of all breast cancers. Although ER alpha positive breast cancer could be effectively controlled by endocrine therapy, more than half of the cases could develop endocrine resistance, making it an important clinical issue in breast cancer treatment. Thus, decoding the detailed mechanism, which controls ER alpha signaling activation and ER alpha protein stability, is of great importance for the improvement of breast cancer therapy. Several zinc finger proteins were shown to mediate the ubiquitination process and modulate protein stability. Thus, we further explore the function of Zinc finger protein 213 on ER alpha protein stability and tamoxifen resistance. METHODS: CCK8 and Edu assay was used to measure cell proliferation. RNA sequence was performed by Ingenuity pathway analysis. The ER alpha signaling activities were measured with luciferase assay, real-time quantitative PCR, and western blotting. Protein stability assay and ubiquitin assay were used to determine ER alpha protein degradation and ubiquitination. The immuno-precipitation was utilized to determine ER alpha and ZNF213 interaction. The ubiquitin-based immuno-precipitation assay was sued to detect specific ubiquitination manner on ER alpha. RESULTS: We identified ZNF213 as a novel zinc finger protein, which modulated ER alpha protein. ZNF213 expression correlated with poor outcome in endocrine treated patients. ZNF213 depletion inhibited ER alpha signaling and proliferation in breast cancer cells. Further mechanistic studies showed ZNF213 located in cytosol and nuclear, which modulated ER alpha stability via inhibiting ER alpha K48-linked ubiquitination. CONCLUSIONS: Our study reveals an interesting post-translational mechanism between ER alpha and ZNF213 in breast cancer. Targeting ZNF213 could be an appealing strategy for ER alpha positive breast cancer.

[Effect of recombinant human growth hormone on the secretion of TNF-α and IL-6 by THP-1 cells and its correlation with the NF-κB pathway].

OBJECTIVE: To investigate the effect of recombinant human growth hormone (rhGH) on the secretion of TNF-α and IL-6 in THP-1 cells and analyze the correlation between the secretion and NF-κB signaling pathway. METHODS: The concentrations of TNF-α and IL-6 in the supernatants of THP-1 induced by rhGH were measured with ELISA. The effects of LPS and an NF-κB inhibitor, BAY11-7082 on the secretion of TNF-α and IL-6 were also observed. The activity of NF-κB was detected by luciferase reporter and electrophoretic mobility shift assay (EMSA) in THP-1 cells induced by rhGH. RESULTS: rhGH alone promoted the secretion of TNF-α and IL-6 in THP-1 cells while inhibited the secretion of TNF-α and IL-6 in THP-1 stimulated by LPS. BAY11-7082 inhibited the secretion of TNF-α and IL-6 in THP-1 stimulated by LPS and rhGH. The activity of NF-κB was significantly correlated with the secretion of TNF-α and IL-6. CONCLUSION: rhGH played a bidirectional role in the regulation on the secretion of TNF-α and IL-6 via NF-κB signaling pathway in THP-1 cells.