Down-regulation of TRIB3 inhibits the progression of ovarian cancer via MEK/ERK signaling pathway

Abstract

Background: Tribbles pseudokinase 3 (TRIB3) protein is a pseudokinase which plays an important role in cellular stress, metabolism, and tumor progression. However, the expression and function of TRIB3 in ovarian cancer is unknown. Methods: TRIB3 expression was detected by immunohistochemistry in the ovarian tissue samples. Following downregulation of TRIB3 by siRNA, multiple aspects of ovarian cancer cells were detected by the MTT assay, flow cytometry, scratch test and Transwell. Additionally, changes in related molecules and the MEK/ERK pathway were detected by western blotting. Finally, many bioinformatic methods, websites and databases, such as gene set enrichment analysis (GSEA), DVAID, Genemania, TISIDB and cBioPortal were used to study the TRIB3. Results: The expression level of TRIB3 was higher in ovarian epithelial malignant tumors as compared to other groups. Patients with a high expression level of TRIB3 had significantly shorter survival times,which was consistent with the results of analysis of the KM-plot database. Down-regulation of TRIB3 expression significantly inhibited the proliferation, invasion, and migration capabilities of ovarian cancer cells, and induced apoptosis and cell cycle arrest. Following TRIB3 siRNA transfection, expression levels of relative proteins were found to be decreased. Additionally, analysis in DAVID website and GSEA revealed that TRIB3 expression was associated with multiple biological processes. Protein phosphorylation levels of MEK and ERK also decreased following TRIB3-siRNA transfection. The Genemania website was used to analyze the proteins that interact with TRIB3. Analysis of TRIB3 in the TISIDB database and cBio-Portal website showed that ovarian cancer patients with high levels of mutation in TRIB3 had poor prognosis, and that the expression of TRIB3 was related to immunomodulation. Conclusions: The TRIB3 was highly expressed and promoting the malignant behavior of ovarian cancer cells by activating the MEK-ERK signaling pathway. It was also found to be associated with genetic variations and immune modulators.

IBSP, a potential recurrence biomarker, promotes the progression of colorectal cancer via Fyn/β‐catenin signaling pathway

“…Western blotting was conducted in accordance with specific protocols. 21 In the present work, the primary antibodies utilized included anti-MMP9 (Proteintech, Cat# 10375-2-AP), anti-PCNA (CST, Cat# 2586S), anti-IBSP (CST, Cat# 5468S), anti-E-cadherin (CST, Cat# 3195T), anti-MMP2 (Proteintech, Cat# 10373-2-AP), anti-N-cadherin (CST, Cat#13116T), anti-Vimentin (CST, Cat# 5741S), anti-CyclinD1 (CST, Cat# 55506S), anti-Cdk4 (CST, Cat# 12790T), anti-Bcl-2 (CST, Cat# 15071T), anti-Bax (CST, Cat# 5023S), anti-Fyn (Abcam, Cat# ab125016), antip-Fyn (phosphor Y530, Abcam, Cat# ab188319), anti-βcatenin (CST, Cat# 8480T), anti-p-β-catenin (phosphor Y142, Abcam, Cat# ab27798), and anti-β-actin (Beyotime Bio Inc, Cat#AA128). Besides, the enhanced chemiluminescence (ECL) detection system was employed to detect the protein-antibody complex.…”

TRIB3 Promotes the Malignant Progression of Bladder Cancer: An Integrated Analysis of Bioinformatics and in vitro Experiments

“…Flow cytometry analysis revealed that arrested G0/G1 cell cycle could be induced by TRIB3-siRNA, confirmed by statistical analysis (Figure 7D), thereby inhibiting the proliferation and migration of bladder cancer cells. Other studies confirmed that the expression of TRIB3 is greater in solid tumors such as colorectal cancer, breast cancer, lung cancer, ovarian cancer, melanoma, liver cancer, and leukemia (Wennemers et al, 2011;Salazar et al, 2015;Hua et al, 2019;Wang et al, 2020;Zhu et al, 2020), suggesting that TRIB3 is a potential target for cancer treatment. Some studies have shown that biological scaffold coordination complexes can be used to design new target-specific therapeutic agents, and TRIB3 is also a scaffold protein that acts as a platform response.…”

“…Studies confirmed that inhibition of the expression of TRIB3 in ovarian cancer cells causes cell cycle arrest in the G0/G1 phase in ovarian cancer cells. Proliferation, migration, and invasion of ovarian cancer cells reduce, and apoptosis increases ( Wang et al, 2020 ). Enrichment analysis of co-expressed genes of TRIB3 showed that TRIB3 was related to the cell cycle ( Figure 6A ).…”

“…These findings suggest that TRIB3 participates in various physiological and pathological processes. The expression of TRIB3 is higher in breast cancer ( Wennemers et al, 2011 ), ovarian cancer ( Wang et al, 2020 ), lung cancer ( Zhang X. et al, 2019 ), and colorectal cancer ( Hua et al, 2019 ), and outcomes are poor. However, few studies reported the role of TRIB3 expression in bladder cancer.…”

Tribbles Pseudokinases in Colorectal Cancer

“…These data support a scenario in which the balance between the expression of Tribbles might determine tumor progression, in this particular case through FOXO proteins activity via AKT. Several studies have been published reporting the role of Tribbles proteins in different types of cancer, including TRIB1 in prostate [ 34 ], glioma [ 35 ], ovarian [ 36 ], and thyroid [ 37 ] cancers; TRIB2 in melanoma [ 38 ], lung [ 39 ], liver [ 40 , 41 , 42 ], acute leukemias [ 27 , 43 , 44 ], and glioblastoma [ 45 ]; and TRIB3 in lung [ 46 ], breast [ 47 , 48 ], renal [ 49 ], gastric [ 50 ], liver [ 51 ], retinoblastoma [ 52 ], glioblastoma [ 53 ], and ovarian [ 54 ] cancers; as well as all Tribbles members in colorectal cancer, as herein fully presented. Tribbles pseudokinases may have an immediate impact in cancer treatment being potential therapeutic targets.…”